Herbert Herscowitz | Georgetown University, School of Medicine (original) (raw)
Papers by Herbert Herscowitz
The Journal of Immunology
Suspensions of popliteal lymph node cells from rabbits primed with keyhole limpet hemocyanin (KLH... more Suspensions of popliteal lymph node cells from rabbits primed with keyhole limpet hemocyanin (KLH) were stimulated in vitro with KLH to give an anamnestic response. Newly synthesized IgG antibody as determined by the incorporation of radioactive leucine was first detected in the culture medium on days 2 and 3, reached a peak on days 5 to 7 and declined thereafter. The antigen-stimulated cells also synthesized increased amounts of non-antibody proteins, including α, β and γ globulins, which accounted for over 90% of the incorporated radioactivity. Both antibody and protein syntheses were antigen dose-dependent and required contact of primed cells with KLH for several hours. The synthesis of DNA and RNA increased in antigen-stimulated cells and preceded the appearance of radioactive antibody.
The Journal of Immunology
Heterologous antisera were prepared against a subpopulation of MOPC-104E tumor cells obtained by ... more Heterologous antisera were prepared against a subpopulation of MOPC-104E tumor cells obtained by centrifugation on discontinuous BSA gradients as well as against cells from the whole tumor mass. The gradient-separated cells were more effective than the cells from the whole tumor mass in eliciting antisera not only of higher titer, but also with greater specificity for plasmacytoma antigens. The unabsorbed antiserum prepared against the gradient-separated plasmacytoma population was cytotoxic for murine lymphoid cells, but not for murine kidney, liver, or brain cells. After in vitro absorption with murine thymocytes and removal of anti-immunoglobulin activity by affinity chromatography, the antiserum was found to be reactive against plasmacytoma cells, but was no longer cytotoxic for murine thymus or unstimulated spleen cells. This absorbed antiserum was also cytotoxic for LPS-, but not PHA- or Con A-stimulated normal murine spleen cells.
Regional immunology, 1994
Infection and Immunity, 1982
Experiments were carried out to determine the effect of cocultivation of T-cell-enriched human pe... more Experiments were carried out to determine the effect of cocultivation of T-cell-enriched human peripheral blood lymphocytes with autologous alveolar macrophages on mitogen-induced proliferation as determined by [ 3 H]thymidine uptake. Cells obtained by fiberoptic bronchoscopy and saline bronchial lavage from 14 normal volunteers were enriched for macrophages by adherence in plastic dishes for 1 h in RPMI 1640 medium supplemented with 10% fetal calf serum. Nonadherent mononuclear cells were prepared from heparinized venous blood after Ficoll-Hypaque sedimentation by passage over nylon wool columns. T-cell-enriched populations were incubated with and without alveolar macrophages, either in the presence or absence of phytohemagglutinin. In these experiments, the number of lymphocytes was held constant (10 5 per well), while the number of alveolar macrophages was varied (0.1 × 10 5 to 4.0 × 10 5 per well). Alveolar macrophages generally tended to stimulate phytohemagglutinin-induced lym...
Infection and Immunity, 1986
The kinetics of induction of the bronchoalveolar cell population (i.e., alveolar macrophages [AM]... more The kinetics of induction of the bronchoalveolar cell population (i.e., alveolar macrophages [AM], lymphocytes, and polymorphonuclear leukocytes) was studied in mice inoculated intravenously with heat-killed Mycobacterium bovis BCG. Injection of BCG at 100 and 500 micrograms but not at 10 micrograms per mouse resulted in an increase in the total number of bronchoalveolar cells (threefold) and in the number of AM (sixfold) recovered by bronchoalveolar lavage in a time-dependent manner, as compared with control mice. A significant increase in the number of lymphocytes was also observed between days 2 and 4 after injection, but this number returned to normal levels by day 8, whereas the number of polymorphonuclear leukocytes was not significantly altered. AM were characteristically phagocytic and stained positively for nonspecific esterase. AM recruited in response to BCG injection were activated, as indicated by elevated levels of acid phosphatase activity and decreased levels of memb...
Journal of Leukocyte Biology, 1984
Rabbit alveolar macrophages (AM) were separated into four subpopulations by centrifugation on dis... more Rabbit alveolar macrophages (AM) were separated into four subpopulations by centrifugation on discontinuous density gradients of Percoll. The subpopulation s were compared to unseparated AM populations for their ability to provide accessory function to ad herent cell-depleted splenocytes for antigen-stimulated Iymphoproliferation and for the production of Iymphokine. They were also tested for their ability to modulate in vitro plaque-forming (PFC) responses. AM subpopulations that provided accessory function for the production of migration inhibitory factor (M IF)-con taini ng cult ure supernantants were recovered from the least dense fractions of the Percoll gradients. These cells were cytoc hemically characterized as mature cells. AM that suppressed the in vitro PFC response and augmented th e antigen-stimulated Iymphoproliferative response to the greatest degree were recovered from the most dense fractions of the Percoll gradients and were characterized as immature cells. These results suggest that there are distinct subpopulatins of AM , the function of which may represe nt different stages of maturation (or differentiation).
Journal of Leukocyte Biology, 1995
We have previously reported that MH-S, an established murine alveolar macrophage-derived cell lin... more We have previously reported that MH-S, an established murine alveolar macrophage-derived cell line, mediated profound inhibition of in vitro antibody production, as did their freshly isolated alveolar macro phage (AM) counterparts. In this communication we show that like freshly recovered AMs, the MH-S cell line also displays phenotypic and functional heterogeneity. Sorting of parental MH-S cells by flow cytometry based on reactivity with anti-Mac-1 antibody yielded two sub sets. Further analysis by staining with monoclonal anti bodies against well-characterized murine macrophage cell surface markers revealed that both Mac-1+ and Mac-1-subsets expressed the mature murine macrophage antigen (F4/80) and class II major histocompatibility complex molecules, but with different intensity. In contrast, the two subsets stained equivalently with antibody against the FCγII receptor, whereas neither subset stained with anti-CD4 antibody. Examination by light microscopy revealed pleomorphism in...
A continuous cell line of murine alveolar macrophages (AM), designated MH-S, has been established... more A continuous cell line of murine alveolar macrophages (AM), designated MH-S, has been establishedfollowingtransformationof cells obtainedby bronchoalveolarlavagefrom BaIb/cJ mice with simian virus 40 (SV4O). Thirty days after infection of the AM cultures, foci of rapidly proliferating cells were recovered and these have been propagated con-tinuously for more than 36 me. Following its initial isolation in Fischer’s medium sup-plemented with L-cell-conditioned medium and horse and fetal bovine serum, the cell line is now routinely grown in RPMI-1640 medium containing 10 % fetal bovine serum in the absence of conditioned medium. MH-S cells were adherent, lacked contact inhibition, and were trypsin-sensitive. They expressed intracellular 1-antigen and incorporated 3H- thy-midine (DNA synthesis) with a doubling time of approximately 48 h but doubled in num-ber in 96 h. MH-S exhibited typical macrophage morphology, was>98 % esterase-posi-tive, negative for peroxidase, and expressed cell surface Ia and Mac-i antigens. The cells were Fc receptor-positive as demonstrated by rosette formation with, and phagocytosis of, antibody-coated sheep erythrocytes. Constitutive IL-i secretion was significantly in-creased following stimulation of the cells with iipopolysaccharlde. Like freshly isolated AM, MH-S cells suppressed the in vitro plaque-forming cell (PFC) response in a dose-dependent manner when cultured with spienic lymphocytes. This cell line should facili-tate studies where homogeneous populations of AM are desirable, especially those in-volved in determining the immunological functions of AM and their potential role in lung pathology. Key words: established cell line, SV4O-transformed, functions
Biochimica et Biophysica Acta (BBA) - Protein Structure, 1970
BBA 33Z89 Pronase and "immunogenic RNA". Inactivation of mouse anti.phage antibody with pronase T... more BBA 33Z89 Pronase and "immunogenic RNA". Inactivation of mouse anti.phage antibody with pronase The induction of an in vitro imnmne response by RNA derived from cells of immune animals or cells ilnmunized iu vitro has been reportedL Such RNA preparations, termed "immunogenic RNA", do not lose their ability to induce antibody formation when treated with deoxyribonuclease, trypsin, or amylase". Digestion with pronase a or ribonucleasO, however, abolishes the imnmnogenicity of these preparations. \Ve have studied RNA extracted from spleens of inmmne mice and found that it initiates synthesis of bacteriophage-neutralizing antibody when added to cultures ~f spleela fragments obtained from nonimmune mice. In our system, the loss of neutral-i×ing antibody activity after pronase treatment of inmmnogenie RNA can be attri-Imted to tile action of ttlis enz\'rne on the neutralizing antibod\' s\nthesized rattier than its action on the RNA extract. Female Swiss mice were injected intraperitoneally with z. IO It plaque forlning units (p.f.u.)of a phage infi>cting .~trc]Stococcus faccalis var. zymo,.gem's on alternate HERBERT 13. HERSCOWITZ* PI-TER STEI.OS
Cellular Immunology, 1988
Studies from this laboratory have demonstrated that incubation of murine alveolar macrophages (AM... more Studies from this laboratory have demonstrated that incubation of murine alveolar macrophages (AM) with SRBC-primed spleen cells (SC) results in suppression of the in vitro plaqueforming cell (PFC) response and that suppression is mediated by a soluble factor contained in supematants obtained from cultures of AM and SC. In the present study, immunological techniques employing monoclonal antibody (MoAb) were used to isolate various T-cell subsets in order to determine the phenotype ofthe cells which interact with AM to produce suppression. Spleen cell populations depleted of Thy-l+-, Lyt-l+-, L3T4+-, or I-J+-bearing cells failed to generate suppressive supematants when cultured with AM. Depletion of Lyt-2+ T-cells (the classical suppressor/effecter subset) did not alter the ability of the remaining cell population to cooperate with AM for generation of suppressive supematants. Direct suppression of the PFC response in cultures containing AM was abrogated after treatment of the spleen cells with anti-I-J, but not anti-Lyt-2 MoAbs. Reconstitution ofthe AM-mediated suppressive response with enriched pop ulations of SC required the presence of T-cells which expressed Lyt-1, L3T4, and I-J. These results suggest the existence of an unusual suppressor pathway involving I-J restriction but which appears to be mediated by the interaction of AM with a population of T-cells that expresses surface markers characteristic of T-helper cells. Q 1988 Academic PXS, Inc.
Journal of the Reticuloendothelial Society, 1979
Methods in Enzymology, 1984
Publisher Summary This chapter describes techniques for the collection of macrophages from the lu... more Publisher Summary This chapter describes techniques for the collection of macrophages from the lungs and peritoneal cavity of rabbits and mice. These basic techniques may be modified, with little effort, to obtain similar cell populations from other species. The collection of alveolar macrophages from rats, dogs, guinea pigs, and humans, as well as the collection of peritoneal macrophages from rats and guinea pigs have also been discussed. The chapter reviews that in order to increase the yield of macrophages obtained from a single animal, an eliciting agent may be injected into the animal prior to the collection of cells. The use of eliciting agents also induces a change in the properties of the collected population as compared to the cells obtained from an untreated animal. It discusses that the techniques for obtaining an elicited population as well as some of the considerations involved in the use of these agents.
Journal of the Reticuloendothelial Society, 1980
Regional immunology
It is known that murine alveolar macrophages function inefficiently as antigen-presenting cells f... more It is known that murine alveolar macrophages function inefficiently as antigen-presenting cells for the in vitro activation of macrophage-depleted T-lymphocyte populations and that they suppress antigen-stimulated lymphoproliferative responses in a dose-dependent manner. The present studies were carried out to determine whether oxidation of the alveolar macrophage surface could alter these activities. Viable alveolar macrophages were treated with varying concentrations of sodium periodate then cultured with either unfractionated or adherent cell-depleted lymphocyte populations obtained from BCG-immunized animals and challenged with PPD. It was demonstrated that moderate oxidation of the alveolar macrophage surface with sodium periodate abrogated their ability to suppress antigen-stimulated proliferation of unfractionated lymphocyte populations, as well as resulted in initiation of lymphoproliferation in an antigen-stimulated, adherent cell-depleted spleen cell populations. It is con...
Prostaglandins, 1975
The ability of various prostaglandins (PGs) to affect the in vitro anamnestic immune response of ... more The ability of various prostaglandins (PGs) to affect the in vitro anamnestic immune response of keyhole limpet hemocyanin (KLH)-primed rabbit popliteal lymph node cells was investigated. Of the four PGs studied (PGAI, PGE 2 and PGF2~) , PGE I was found to have a stimulatory effect, whereas PGAI, PGE 2 and PGF2~ were ineffective in stimulating or inhibiting the in vitro anamnestic response. Under the conditions studied, a 3.5-fold increase in antibody production was obtained in PGEl-treated , KLH-stimulated cultures. Maximum enhancement was obtained when 0.2 ~g of PGE I were added at the time of culture initiation and were allowed to remain in contact with the lymph node cells for 24 hours.
Journal of Hematotherapy & Stem Cell Research, 2001
Ex vivo activation of peripheral blood stem cells (PBSC) using interleukin-2 (IL-2) results in cy... more Ex vivo activation of peripheral blood stem cells (PBSC) using interleukin-2 (IL-2) results in cytotoxic effector cells that may possess beneficial in vivo effects. We proposed to evaluate ex vivo stimulation of PBSC using various cytokines alone or in combination to optimize their function. Cytokine-activated PBSC were analyzed for tumor-directed cytotoxicity and their ability to remove tumor cells from long-term clonogenic assays. Mononuclear cells were obtained from the apheresis products of normal donors and cultured with IL-2 (1000 U/ml), interferon-alpha (IFN-alpha) (1000 U/ml), or IL-12 (50 U/ml) either alone or in combinations at 37 degrees C and 5% CO(2) for 24 h. Colony-forming unit-tumor (CFUT) assays were initiated using cytokine-activated PBSC with varying concentrations of MCF-7 or SKBR-3 human breast cancer cells. Standard 4-h (51)Cr-release assays were performed with cytokine-activated PBSC using MCF-7 or SKBR-3 cells as targets. Activation of PBSC with IL-2, IFN-alpha, or IL-12 resulted in enhanced cytotoxicity against the two breast cancer cell lines when compared to controls. PBSC activated with IL-2 and IFN-alpha or IL-2 and IL-12 were more cytotoxic than PBSC activated with single cytokines (p = 0.0004 for MCF-7 cells and p < 0.001 for SKBR-3 cells). Using clonogenic assays, IL-2-activated PBSC reduced the number of CFU-T to a greater extent than did IL-12 or IFN-alpha-activated PBSC (p = 0.0006). However, PBSC activated with a combination of IL-2 and IFN-alpha or IL-2 and IL-12 demonstrated 95% and 90% reductions, respectively, compared to 79% reduction using IL-2-activated PBSC (p < 0.0001). The greatest reduction in cytotoxicity occurred in the cell populations depleted of CD56(+) cells (p = 0.016) and CD8(+) CD56(+) cells (p = 0.002), suggesting that the effector cell population includes a combination of cytotoxic CD8(+) T cells and CD56(+) natural killer cells. These results demonstrate that the ex vivo activation of PBSC with cytokines, either alone or in combination, enhances cytotoxicity against, and removal of two human breast cancer cells. The combinations of IL-2 with IFN-alpha or IL-12 are most beneficial in cytotoxicity and purging assays. These results could play an important role in designing adoptive cellular immunotherapy clinical trials in the autologous hematopoietic stem cell transplant setting.
Journal of Hematotherapy & Stem Cell Research, 2003
Effective mobilization of peripheral blood stem cells is vital for transplantation of patients af... more Effective mobilization of peripheral blood stem cells is vital for transplantation of patients after high-dose chemotherapy and provides a convenient source of stem cells for genetic engineering and other studies, but optimal mobilization strategies have not been defined. Recent studies show that in the presence of recombinant human granulocyte colony-stimulating factor (rhG-CSF), docetaxel (DXT) is an effective mobilization agent. This study was performed to evaluate the phenotype and immunologic properties of DXT-mobilized stem cells. Administration of DXT + rhG-CSF to normal C57Bl/6 mice induced a 75-fold increase in blood hematopoietic progenitors and a significant increase in both CD3(+) (T cell) and DX5(+) [natural killer (NK)] cells when compared to untreated mice. The cytotoxicity of DXT + rhG-CSF-mobilized cell populations against YAC-1 and B16F10 cell lines was not significantly different from that of untreated mice. When compared to cyclophosphamide + rhG-CSF, DXT + rhG-CSF-mobilized cell populations yielded a greater number of T and NK cells, with significantly higher cytotoxic effector function. These results suggest that DXT + rhG-CSF-mobilized PBSCs retain potent immunologic capacity with a high number of the functional cellular subsets than those normally present in peripheral blood, which may be important in maintaining the antitumor immunity after transplantation.
Journal of Medical Microbiology, 1982
circumspection. The reason that batteries of short-term tests are recommended by regulatory autho... more circumspection. The reason that batteries of short-term tests are recommended by regulatory authorities is very apparent. On the theoretical side, the close correlation between DNA damage and carcinogenesis further strengthens the case for the somatic mutation hypothesis of tumour initiation. The book is well produced and contains few errors. However, the reviewer (a chemist) takes exception to the description, on p. 97, of 4-nitro-quinoline oxide as a "water-soluble hydrocarbon". M. M. COOMBS The biochemistry and pharmacology of antibacterial agents By R. A. D. WILLIAMS and Z. L. KRUK. 1981. Croom Helm Ltd, London. Pp. 89. E3.95. This is a modest little book, hardly more than a long article, in which the barest skeleton of an account of the biochemistry and pharmacology of antibacterial agents is laid out. Within its compass the text is clear and simple and its coverage is really the minimum of what many in medicine and paramedical subjects might be expected to know about the molecular basis of how antibacterial agents work. The subject is tackled from the point of view of the targets that antibacterial agents attack in the microbial cell. There are sections on agents active against the folic acid pathway, peptidoglycan biosynthesis, protein synthesis and nucleic acid synthesis, but one lacks a chapter on agents active on membranes. In particular, I find no consolidated account of how chemical molecules interact with enzymes. So the fundamental molecular basis
Journal of Immunological Methods, 1974
A comparison of conventional gamma spectrometer and liquid scintillation counting methods was mad... more A comparison of conventional gamma spectrometer and liquid scintillation counting methods was made using gamma-emitting isotopes employed in studies of antigen-antibody interactions and cell-mediated immunity. Results indicate that 12SIodine and 51Chromium can be counted with equal, if not greater efficiency by a liquid scintillation detector than with a conventional gamma spectrometer. The method described is rapid, inexpensive, sensitive and has wide application in research and clinical laboratories.
Journal of Immunological Methods, 1987
A monoclonal antibody prepared against the murine interleukin-2 receptor (IL-2R) was employed to ... more A monoclonal antibody prepared against the murine interleukin-2 receptor (IL-2R) was employed to develop an ELISA method for measuring the immunological activation of T-cells. The assay detects an increase in IL-2R expression on activated lymphocytes. Stimulated splenic lymphocytes displayed markedly higher IL-2R expression compared to unstimulated controls. A significant increase in IL-2R expression on lymphocytes was detected in mitogen-stimulated responses, in a one-way mixed leukocyte reaction (MLR) and in the antigen-specific responses to conalbumin and purified protein derivative (PPD) in vitro. At a constant cell number, the level of IL-2R expression was found to be dependent on the dose of the stimulant. A comparative study of the kinetics of activation of splenic lymphocytes in response to mitogen, antigen and allogeneic cells as measured by the IL-2R ELISA and the conventional tritiated thymidine (3HTdR) uptake assay revealed remarkable similarity. For both assays, the mitogenic response was detected within 12 h and peaked at 72 h, the MLR was detectable within 2-3 days and peaked at day 6, and the specific antigenic response was detected within 2 days and peaked on day 4-5. Hydroxyurea, an inhibitor of DNA synthesis, had no effect on early IL-2R expression by mitogen-stimulated splenic lymphocytes, however, only 20% of maximum IL-2R expression could be detected at later stages of incubation. In contrast, cycloheximide, an inhibitor of protein synthesis, completely abrogated IL-2R expression and proliferation of stimulated lymphocytes.
The Journal of Immunology
Suspensions of popliteal lymph node cells from rabbits primed with keyhole limpet hemocyanin (KLH... more Suspensions of popliteal lymph node cells from rabbits primed with keyhole limpet hemocyanin (KLH) were stimulated in vitro with KLH to give an anamnestic response. Newly synthesized IgG antibody as determined by the incorporation of radioactive leucine was first detected in the culture medium on days 2 and 3, reached a peak on days 5 to 7 and declined thereafter. The antigen-stimulated cells also synthesized increased amounts of non-antibody proteins, including α, β and γ globulins, which accounted for over 90% of the incorporated radioactivity. Both antibody and protein syntheses were antigen dose-dependent and required contact of primed cells with KLH for several hours. The synthesis of DNA and RNA increased in antigen-stimulated cells and preceded the appearance of radioactive antibody.
The Journal of Immunology
Heterologous antisera were prepared against a subpopulation of MOPC-104E tumor cells obtained by ... more Heterologous antisera were prepared against a subpopulation of MOPC-104E tumor cells obtained by centrifugation on discontinuous BSA gradients as well as against cells from the whole tumor mass. The gradient-separated cells were more effective than the cells from the whole tumor mass in eliciting antisera not only of higher titer, but also with greater specificity for plasmacytoma antigens. The unabsorbed antiserum prepared against the gradient-separated plasmacytoma population was cytotoxic for murine lymphoid cells, but not for murine kidney, liver, or brain cells. After in vitro absorption with murine thymocytes and removal of anti-immunoglobulin activity by affinity chromatography, the antiserum was found to be reactive against plasmacytoma cells, but was no longer cytotoxic for murine thymus or unstimulated spleen cells. This absorbed antiserum was also cytotoxic for LPS-, but not PHA- or Con A-stimulated normal murine spleen cells.
Regional immunology, 1994
Infection and Immunity, 1982
Experiments were carried out to determine the effect of cocultivation of T-cell-enriched human pe... more Experiments were carried out to determine the effect of cocultivation of T-cell-enriched human peripheral blood lymphocytes with autologous alveolar macrophages on mitogen-induced proliferation as determined by [ 3 H]thymidine uptake. Cells obtained by fiberoptic bronchoscopy and saline bronchial lavage from 14 normal volunteers were enriched for macrophages by adherence in plastic dishes for 1 h in RPMI 1640 medium supplemented with 10% fetal calf serum. Nonadherent mononuclear cells were prepared from heparinized venous blood after Ficoll-Hypaque sedimentation by passage over nylon wool columns. T-cell-enriched populations were incubated with and without alveolar macrophages, either in the presence or absence of phytohemagglutinin. In these experiments, the number of lymphocytes was held constant (10 5 per well), while the number of alveolar macrophages was varied (0.1 × 10 5 to 4.0 × 10 5 per well). Alveolar macrophages generally tended to stimulate phytohemagglutinin-induced lym...
Infection and Immunity, 1986
The kinetics of induction of the bronchoalveolar cell population (i.e., alveolar macrophages [AM]... more The kinetics of induction of the bronchoalveolar cell population (i.e., alveolar macrophages [AM], lymphocytes, and polymorphonuclear leukocytes) was studied in mice inoculated intravenously with heat-killed Mycobacterium bovis BCG. Injection of BCG at 100 and 500 micrograms but not at 10 micrograms per mouse resulted in an increase in the total number of bronchoalveolar cells (threefold) and in the number of AM (sixfold) recovered by bronchoalveolar lavage in a time-dependent manner, as compared with control mice. A significant increase in the number of lymphocytes was also observed between days 2 and 4 after injection, but this number returned to normal levels by day 8, whereas the number of polymorphonuclear leukocytes was not significantly altered. AM were characteristically phagocytic and stained positively for nonspecific esterase. AM recruited in response to BCG injection were activated, as indicated by elevated levels of acid phosphatase activity and decreased levels of memb...
Journal of Leukocyte Biology, 1984
Rabbit alveolar macrophages (AM) were separated into four subpopulations by centrifugation on dis... more Rabbit alveolar macrophages (AM) were separated into four subpopulations by centrifugation on discontinuous density gradients of Percoll. The subpopulation s were compared to unseparated AM populations for their ability to provide accessory function to ad herent cell-depleted splenocytes for antigen-stimulated Iymphoproliferation and for the production of Iymphokine. They were also tested for their ability to modulate in vitro plaque-forming (PFC) responses. AM subpopulations that provided accessory function for the production of migration inhibitory factor (M IF)-con taini ng cult ure supernantants were recovered from the least dense fractions of the Percoll gradients. These cells were cytoc hemically characterized as mature cells. AM that suppressed the in vitro PFC response and augmented th e antigen-stimulated Iymphoproliferative response to the greatest degree were recovered from the most dense fractions of the Percoll gradients and were characterized as immature cells. These results suggest that there are distinct subpopulatins of AM , the function of which may represe nt different stages of maturation (or differentiation).
Journal of Leukocyte Biology, 1995
We have previously reported that MH-S, an established murine alveolar macrophage-derived cell lin... more We have previously reported that MH-S, an established murine alveolar macrophage-derived cell line, mediated profound inhibition of in vitro antibody production, as did their freshly isolated alveolar macro phage (AM) counterparts. In this communication we show that like freshly recovered AMs, the MH-S cell line also displays phenotypic and functional heterogeneity. Sorting of parental MH-S cells by flow cytometry based on reactivity with anti-Mac-1 antibody yielded two sub sets. Further analysis by staining with monoclonal anti bodies against well-characterized murine macrophage cell surface markers revealed that both Mac-1+ and Mac-1-subsets expressed the mature murine macrophage antigen (F4/80) and class II major histocompatibility complex molecules, but with different intensity. In contrast, the two subsets stained equivalently with antibody against the FCγII receptor, whereas neither subset stained with anti-CD4 antibody. Examination by light microscopy revealed pleomorphism in...
A continuous cell line of murine alveolar macrophages (AM), designated MH-S, has been established... more A continuous cell line of murine alveolar macrophages (AM), designated MH-S, has been establishedfollowingtransformationof cells obtainedby bronchoalveolarlavagefrom BaIb/cJ mice with simian virus 40 (SV4O). Thirty days after infection of the AM cultures, foci of rapidly proliferating cells were recovered and these have been propagated con-tinuously for more than 36 me. Following its initial isolation in Fischer’s medium sup-plemented with L-cell-conditioned medium and horse and fetal bovine serum, the cell line is now routinely grown in RPMI-1640 medium containing 10 % fetal bovine serum in the absence of conditioned medium. MH-S cells were adherent, lacked contact inhibition, and were trypsin-sensitive. They expressed intracellular 1-antigen and incorporated 3H- thy-midine (DNA synthesis) with a doubling time of approximately 48 h but doubled in num-ber in 96 h. MH-S exhibited typical macrophage morphology, was>98 % esterase-posi-tive, negative for peroxidase, and expressed cell surface Ia and Mac-i antigens. The cells were Fc receptor-positive as demonstrated by rosette formation with, and phagocytosis of, antibody-coated sheep erythrocytes. Constitutive IL-i secretion was significantly in-creased following stimulation of the cells with iipopolysaccharlde. Like freshly isolated AM, MH-S cells suppressed the in vitro plaque-forming cell (PFC) response in a dose-dependent manner when cultured with spienic lymphocytes. This cell line should facili-tate studies where homogeneous populations of AM are desirable, especially those in-volved in determining the immunological functions of AM and their potential role in lung pathology. Key words: established cell line, SV4O-transformed, functions
Biochimica et Biophysica Acta (BBA) - Protein Structure, 1970
BBA 33Z89 Pronase and "immunogenic RNA". Inactivation of mouse anti.phage antibody with pronase T... more BBA 33Z89 Pronase and "immunogenic RNA". Inactivation of mouse anti.phage antibody with pronase The induction of an in vitro imnmne response by RNA derived from cells of immune animals or cells ilnmunized iu vitro has been reportedL Such RNA preparations, termed "immunogenic RNA", do not lose their ability to induce antibody formation when treated with deoxyribonuclease, trypsin, or amylase". Digestion with pronase a or ribonucleasO, however, abolishes the imnmnogenicity of these preparations. \Ve have studied RNA extracted from spleens of inmmne mice and found that it initiates synthesis of bacteriophage-neutralizing antibody when added to cultures ~f spleela fragments obtained from nonimmune mice. In our system, the loss of neutral-i×ing antibody activity after pronase treatment of inmmnogenie RNA can be attri-Imted to tile action of ttlis enz\'rne on the neutralizing antibod\' s\nthesized rattier than its action on the RNA extract. Female Swiss mice were injected intraperitoneally with z. IO It plaque forlning units (p.f.u.)of a phage infi>cting .~trc]Stococcus faccalis var. zymo,.gem's on alternate HERBERT 13. HERSCOWITZ* PI-TER STEI.OS
Cellular Immunology, 1988
Studies from this laboratory have demonstrated that incubation of murine alveolar macrophages (AM... more Studies from this laboratory have demonstrated that incubation of murine alveolar macrophages (AM) with SRBC-primed spleen cells (SC) results in suppression of the in vitro plaqueforming cell (PFC) response and that suppression is mediated by a soluble factor contained in supematants obtained from cultures of AM and SC. In the present study, immunological techniques employing monoclonal antibody (MoAb) were used to isolate various T-cell subsets in order to determine the phenotype ofthe cells which interact with AM to produce suppression. Spleen cell populations depleted of Thy-l+-, Lyt-l+-, L3T4+-, or I-J+-bearing cells failed to generate suppressive supematants when cultured with AM. Depletion of Lyt-2+ T-cells (the classical suppressor/effecter subset) did not alter the ability of the remaining cell population to cooperate with AM for generation of suppressive supematants. Direct suppression of the PFC response in cultures containing AM was abrogated after treatment of the spleen cells with anti-I-J, but not anti-Lyt-2 MoAbs. Reconstitution ofthe AM-mediated suppressive response with enriched pop ulations of SC required the presence of T-cells which expressed Lyt-1, L3T4, and I-J. These results suggest the existence of an unusual suppressor pathway involving I-J restriction but which appears to be mediated by the interaction of AM with a population of T-cells that expresses surface markers characteristic of T-helper cells. Q 1988 Academic PXS, Inc.
Journal of the Reticuloendothelial Society, 1979
Methods in Enzymology, 1984
Publisher Summary This chapter describes techniques for the collection of macrophages from the lu... more Publisher Summary This chapter describes techniques for the collection of macrophages from the lungs and peritoneal cavity of rabbits and mice. These basic techniques may be modified, with little effort, to obtain similar cell populations from other species. The collection of alveolar macrophages from rats, dogs, guinea pigs, and humans, as well as the collection of peritoneal macrophages from rats and guinea pigs have also been discussed. The chapter reviews that in order to increase the yield of macrophages obtained from a single animal, an eliciting agent may be injected into the animal prior to the collection of cells. The use of eliciting agents also induces a change in the properties of the collected population as compared to the cells obtained from an untreated animal. It discusses that the techniques for obtaining an elicited population as well as some of the considerations involved in the use of these agents.
Journal of the Reticuloendothelial Society, 1980
Regional immunology
It is known that murine alveolar macrophages function inefficiently as antigen-presenting cells f... more It is known that murine alveolar macrophages function inefficiently as antigen-presenting cells for the in vitro activation of macrophage-depleted T-lymphocyte populations and that they suppress antigen-stimulated lymphoproliferative responses in a dose-dependent manner. The present studies were carried out to determine whether oxidation of the alveolar macrophage surface could alter these activities. Viable alveolar macrophages were treated with varying concentrations of sodium periodate then cultured with either unfractionated or adherent cell-depleted lymphocyte populations obtained from BCG-immunized animals and challenged with PPD. It was demonstrated that moderate oxidation of the alveolar macrophage surface with sodium periodate abrogated their ability to suppress antigen-stimulated proliferation of unfractionated lymphocyte populations, as well as resulted in initiation of lymphoproliferation in an antigen-stimulated, adherent cell-depleted spleen cell populations. It is con...
Prostaglandins, 1975
The ability of various prostaglandins (PGs) to affect the in vitro anamnestic immune response of ... more The ability of various prostaglandins (PGs) to affect the in vitro anamnestic immune response of keyhole limpet hemocyanin (KLH)-primed rabbit popliteal lymph node cells was investigated. Of the four PGs studied (PGAI, PGE 2 and PGF2~) , PGE I was found to have a stimulatory effect, whereas PGAI, PGE 2 and PGF2~ were ineffective in stimulating or inhibiting the in vitro anamnestic response. Under the conditions studied, a 3.5-fold increase in antibody production was obtained in PGEl-treated , KLH-stimulated cultures. Maximum enhancement was obtained when 0.2 ~g of PGE I were added at the time of culture initiation and were allowed to remain in contact with the lymph node cells for 24 hours.
Journal of Hematotherapy & Stem Cell Research, 2001
Ex vivo activation of peripheral blood stem cells (PBSC) using interleukin-2 (IL-2) results in cy... more Ex vivo activation of peripheral blood stem cells (PBSC) using interleukin-2 (IL-2) results in cytotoxic effector cells that may possess beneficial in vivo effects. We proposed to evaluate ex vivo stimulation of PBSC using various cytokines alone or in combination to optimize their function. Cytokine-activated PBSC were analyzed for tumor-directed cytotoxicity and their ability to remove tumor cells from long-term clonogenic assays. Mononuclear cells were obtained from the apheresis products of normal donors and cultured with IL-2 (1000 U/ml), interferon-alpha (IFN-alpha) (1000 U/ml), or IL-12 (50 U/ml) either alone or in combinations at 37 degrees C and 5% CO(2) for 24 h. Colony-forming unit-tumor (CFUT) assays were initiated using cytokine-activated PBSC with varying concentrations of MCF-7 or SKBR-3 human breast cancer cells. Standard 4-h (51)Cr-release assays were performed with cytokine-activated PBSC using MCF-7 or SKBR-3 cells as targets. Activation of PBSC with IL-2, IFN-alpha, or IL-12 resulted in enhanced cytotoxicity against the two breast cancer cell lines when compared to controls. PBSC activated with IL-2 and IFN-alpha or IL-2 and IL-12 were more cytotoxic than PBSC activated with single cytokines (p = 0.0004 for MCF-7 cells and p < 0.001 for SKBR-3 cells). Using clonogenic assays, IL-2-activated PBSC reduced the number of CFU-T to a greater extent than did IL-12 or IFN-alpha-activated PBSC (p = 0.0006). However, PBSC activated with a combination of IL-2 and IFN-alpha or IL-2 and IL-12 demonstrated 95% and 90% reductions, respectively, compared to 79% reduction using IL-2-activated PBSC (p < 0.0001). The greatest reduction in cytotoxicity occurred in the cell populations depleted of CD56(+) cells (p = 0.016) and CD8(+) CD56(+) cells (p = 0.002), suggesting that the effector cell population includes a combination of cytotoxic CD8(+) T cells and CD56(+) natural killer cells. These results demonstrate that the ex vivo activation of PBSC with cytokines, either alone or in combination, enhances cytotoxicity against, and removal of two human breast cancer cells. The combinations of IL-2 with IFN-alpha or IL-12 are most beneficial in cytotoxicity and purging assays. These results could play an important role in designing adoptive cellular immunotherapy clinical trials in the autologous hematopoietic stem cell transplant setting.
Journal of Hematotherapy & Stem Cell Research, 2003
Effective mobilization of peripheral blood stem cells is vital for transplantation of patients af... more Effective mobilization of peripheral blood stem cells is vital for transplantation of patients after high-dose chemotherapy and provides a convenient source of stem cells for genetic engineering and other studies, but optimal mobilization strategies have not been defined. Recent studies show that in the presence of recombinant human granulocyte colony-stimulating factor (rhG-CSF), docetaxel (DXT) is an effective mobilization agent. This study was performed to evaluate the phenotype and immunologic properties of DXT-mobilized stem cells. Administration of DXT + rhG-CSF to normal C57Bl/6 mice induced a 75-fold increase in blood hematopoietic progenitors and a significant increase in both CD3(+) (T cell) and DX5(+) [natural killer (NK)] cells when compared to untreated mice. The cytotoxicity of DXT + rhG-CSF-mobilized cell populations against YAC-1 and B16F10 cell lines was not significantly different from that of untreated mice. When compared to cyclophosphamide + rhG-CSF, DXT + rhG-CSF-mobilized cell populations yielded a greater number of T and NK cells, with significantly higher cytotoxic effector function. These results suggest that DXT + rhG-CSF-mobilized PBSCs retain potent immunologic capacity with a high number of the functional cellular subsets than those normally present in peripheral blood, which may be important in maintaining the antitumor immunity after transplantation.
Journal of Medical Microbiology, 1982
circumspection. The reason that batteries of short-term tests are recommended by regulatory autho... more circumspection. The reason that batteries of short-term tests are recommended by regulatory authorities is very apparent. On the theoretical side, the close correlation between DNA damage and carcinogenesis further strengthens the case for the somatic mutation hypothesis of tumour initiation. The book is well produced and contains few errors. However, the reviewer (a chemist) takes exception to the description, on p. 97, of 4-nitro-quinoline oxide as a "water-soluble hydrocarbon". M. M. COOMBS The biochemistry and pharmacology of antibacterial agents By R. A. D. WILLIAMS and Z. L. KRUK. 1981. Croom Helm Ltd, London. Pp. 89. E3.95. This is a modest little book, hardly more than a long article, in which the barest skeleton of an account of the biochemistry and pharmacology of antibacterial agents is laid out. Within its compass the text is clear and simple and its coverage is really the minimum of what many in medicine and paramedical subjects might be expected to know about the molecular basis of how antibacterial agents work. The subject is tackled from the point of view of the targets that antibacterial agents attack in the microbial cell. There are sections on agents active against the folic acid pathway, peptidoglycan biosynthesis, protein synthesis and nucleic acid synthesis, but one lacks a chapter on agents active on membranes. In particular, I find no consolidated account of how chemical molecules interact with enzymes. So the fundamental molecular basis
Journal of Immunological Methods, 1974
A comparison of conventional gamma spectrometer and liquid scintillation counting methods was mad... more A comparison of conventional gamma spectrometer and liquid scintillation counting methods was made using gamma-emitting isotopes employed in studies of antigen-antibody interactions and cell-mediated immunity. Results indicate that 12SIodine and 51Chromium can be counted with equal, if not greater efficiency by a liquid scintillation detector than with a conventional gamma spectrometer. The method described is rapid, inexpensive, sensitive and has wide application in research and clinical laboratories.
Journal of Immunological Methods, 1987
A monoclonal antibody prepared against the murine interleukin-2 receptor (IL-2R) was employed to ... more A monoclonal antibody prepared against the murine interleukin-2 receptor (IL-2R) was employed to develop an ELISA method for measuring the immunological activation of T-cells. The assay detects an increase in IL-2R expression on activated lymphocytes. Stimulated splenic lymphocytes displayed markedly higher IL-2R expression compared to unstimulated controls. A significant increase in IL-2R expression on lymphocytes was detected in mitogen-stimulated responses, in a one-way mixed leukocyte reaction (MLR) and in the antigen-specific responses to conalbumin and purified protein derivative (PPD) in vitro. At a constant cell number, the level of IL-2R expression was found to be dependent on the dose of the stimulant. A comparative study of the kinetics of activation of splenic lymphocytes in response to mitogen, antigen and allogeneic cells as measured by the IL-2R ELISA and the conventional tritiated thymidine (3HTdR) uptake assay revealed remarkable similarity. For both assays, the mitogenic response was detected within 12 h and peaked at 72 h, the MLR was detectable within 2-3 days and peaked at day 6, and the specific antigenic response was detected within 2 days and peaked on day 4-5. Hydroxyurea, an inhibitor of DNA synthesis, had no effect on early IL-2R expression by mitogen-stimulated splenic lymphocytes, however, only 20% of maximum IL-2R expression could be detected at later stages of incubation. In contrast, cycloheximide, an inhibitor of protein synthesis, completely abrogated IL-2R expression and proliferation of stimulated lymphocytes.