Anti-tumor antibody produced by human tumor-infiltrating and peripheral blood B lymphocytes (original) (raw)
Related papers
Cancer Science, 2005
We previously demonstrated that TIB recognize tumor antigens and produce antibodies against them. In the present study, we identified three tumor antigens recognized by TIB in lung cancer and evaluated whether changes in the antibody titer against these antigens correlated with the patient's clinical course. A lung cancer cell line, G603L, was established from a primary lung tumor of a patient, G603. Seven months later, adrenal metastasis was detected and surgically resected. The latter tumor was mildly infiltrated with B cells and xenotransplanted into SCID mice to obtain human IgG. A cDNA library was constructed from G603L and SEREX was carried out using TIB-derived IgG. The seroreactive clones were sequenced and one of these antigens was revealed to be MAGE-B2 whereas the others were novel antigens. In the immunomonitoring of the patient's sera, high antibody titer against MAGE-B2 was observed before operation and the titer decreased after resection of the primary tumor. It was elevated again at the time of adrenal metastasis, but then decreased after resection. The change in antibody titer against the second antigen was similar to MAGE-B2, and the antibody titer against the third antigen was low before the primary operation but increased at the time of recurrence. Our results suggest that TIB recognized tumor antigens and the antibody titers against these antigens were changed along with the patient's clinical course. Therefore, these antibodies could be used as tumor markers for the patient. (Cancer Sci 2005; 96: 882-888)
Effect of IgG Produced by Tumor-infiltrating B Lymphocytes on Lung Tumor Growth
Anticancer Research, 2006
Background: Tumor-infiltrating B lymphocytes (TIB) are often observed in lung cancer. The role of TIB in tumor growth has not been well investigated. Materials and Methods: Forty-four surgically-resected human lung cancer tissues were xenotransplanted into SCID mice. Their blood was collected and the volume of the transplanted tumors was measured regularly. The correlations between the IgG titer in the sera and the growth of the transplanted tumors according to the clinicopathological variables were examined. Results: Human IgG production from TIB was observed in the all xenotransplanted mice. Twenty-seven out of the 44 tumors regressed gradually. The average serum human IgG level of the tumor regressors (n=10) was significantly higher than that of the progressors (n=9) in squamous cell carcinoma (p=0.02), while there was no significant difference in the other histological groups. Conclusion: IgG produced by TIB might play a crucial role in preventing tumor growth in squamous cell carcinoma. 1827 Abbreviations: SCID mouse, severe combined immunodeficiency mouse; TIL, tumor-infiltrating lymphocytes; TIB, tumor-infiltrating B lymphocytes; SEREX, serological analysis of recombinant cDNA expression libraries of human tumors. Correspondence to: Makiko Mizukami, MD, Second
BMC Biotechnology, 2007
Background There is much evidence that tumor cells elicit a humoral immune response in patients. In most cases, the presence of antibodies in peripheral blood is detected only in small proportion of patients with tumors overexpressing the corresponding antigen. In the present study, we analyzed the significance of local humoral response provided by tumor-infiltrating lymphocytes in breast cancer patients. Methods The ability of a patient's immune system to produce specific antibodies inside tumor tissue, capable of recognizing tumor cells, was explored through analysis of the oligoclonality of antibodies derived from tumor-infiltrating lymphocytes and construction of a series of recombinant antibody libraries in scFv format, derived from breast tumor-infiltrating B lymphocytes. These libraries and one from peripheral blood lymphocytes of a single breast cancer patient were panned against three purified surface tumor antigens, such as CEA, MUC1 and ED-B domain, and against intact MCF7 breast carcinoma cells. Results Application of novel display vector, pKM19, allowed isolation of a large panel of breast cancer-specific antibodies against known tumor antigens, as well as against breast carcinoma cells. Reactivity of novel scFvs was confirmed by ELISA, immunohistochemistry, fluorescence staining and flow cytometry. We demonstrated that seven of ten primary breast tumor specimens, obtained using discarded surgical material, could be exploited as an appropriate source for generation of phage display libraries, giving highly specific antitumor antibodies which recognize heterologous tumor cells. Conclusion Local humoral immune response within tumor tissue in breast cancer patients frequently has an oligoclonal character. Efficient selection of specific antitumor antibodies from recombinant antibody libraries, derived from such oligoclonal tumor-infiltrated B lymphocytes, indicates the presence of natural immune response against tumor antigens in these patients. The described method is very promising for development of antitumor antibodies, potentially useful for diagnostic and therapeutic approaches.
Analysis of the interaction of monoclonal antibodies with surface IgM on neoplastic B-cells
British Journal of Cancer, 1999
In vitro studies have shown that crosslinking of the B-cell receptor (BCR) by anti-IgM monoclonal antibody (mAb) can have either stimulatory or inhibitory effects depending on the maturation state of the cell (De Franco et al, 1982; Hasbold and Klaus, 1990.). Of particular interest in the field of immunotherapy is that crosslinking of the BCR on B lymphoma cells can induce inhibitory signals that may result in cell cycle arrest and apoptosis (Hasbold and Klaus, 1990; Marches et al, 1995). There is growing evidence that such transmembrane signalling is critical to much of the therapeutic success of anti-idiotype (Id) mAb in the treatment of non-Hodgkins lymphoma (NHL). For example the study by Vuist et al (1994) highlighted the close correlation between the therapeutic success of anti-Id mAb in NHL and the ability of the treatment mAb to stimulate protein tyrosine phosphorylation in frozen archive tumour material. Since tyrosine phosphorylation following crosslinking of surface immunoglobulin (Ig) is one of the first steps in the cascade of intracellular signalling events (Gold et al, 1991; Sefton and Campbell, 1991), this positive correlation suggests that the key to the success of anti-Id mAb may stem from their ability to crosslink surface Ig and deliver a direct anti-proliferative signal to the tumour. The importance of crosslinking was further suggested by the observation that although cells from non-responding patients could not be induced to signal with anti-Id mAb, they were responsive to stimulation with a polyclonal anti-µ Ab, which, because it recognizes multiple epitopes on the BCR, possesses a far greater potential for crosslinking than a single mAb. While these results indicate the importance of crosslinking in the inhibitory action of anti-Id mAb, the extent of crosslinking necessary for achieving inhibition, and the antibody binding requirements, have not been fully defined. In this paper, we describe the sensitivity of a panel of Burkitt's lymphoma cell lines to the growth inhibitory effects of anti-IgM mAb, and a series of experiments designed to look at the effects of IgM domain specificity, BCR crosslinking, and hyper-crosslinking on growth inhibition. Cell lines and antibodies The Burkitt's lymphoma cell lines Daudi, Ramos, Ramos-sublines RA.1 and EHRB, Raji, Namalwa, MUTU-I and MUTU-61E (kindly given by Dr A Milner and Prof. Chris Gregory, Birmingham, UK), Nalm-6 (kindly given by Dr Bill Cushley, Glasgow, UK) and Bristol-8 cells were used in this study. All cell lines were maintained in supplemented RPMI-1640 [RPMI-1640 medium containing glutamine (2 mM), pyruvate (1 mM), penicillin and streptomycin (100 IU ml-1), fungizone (2 µg ml-1) and 10% fetal calf serum (FCS) (Myoclone) (Gibco Ltd, Paisley, UK)]. Monoclonal antibodies used in these studies included: anti-µ, M15/8, ZL7/5 and Mc41/24G10 (all anti-Fcµ), and anti-λ, M15/2 and Mc24/1C6, all raised in this laboratory (Bonardi et al, 1993; Zhang et al, 1995), the anti-Fabµ antibodies M-2E6 (ATCC, Rockville, MA, USA) and XG9 (kindly given by Dr Patricia Mongini, New York, NY, USA) (Rudich et al, 1988). In addition, we used three anti-Id mAb, ZL16/1, ZL16/3 and ZL16/11, specific for the surface IgM of the Ramos cell line and one anti-Id mAb, ZL15/27, specific for the surface IgM of the Daudi cell line (Zhang et al, 1995). Polyclonal sheep anti-mouse IgG was raised at Tenovus. All hybridoma lines secreting mAb were expanded in tissue culture and purified from culture supernatant using a Protein A column (Glennie et al, 1993). F(ab′) 2 fragments of IgG were
Cancer Research
Two new monoclonal antibodies (Lym-1 and Lym-2), reactive with the cell surface of B-lymphocytes and derived tumors, have been produced using tumor cell nuclei preparations as immunogens. Specificity screens using live cell radioimmunoassay techniques with 52 well-characterized human lymphoma and leukemia cell lines showed that both Lym-1 and Lym-2 bound to cell lines of B-cell lineage but were unreactive with those of T-cell, myeloid, or erythroid derivation. The B-cell specificity of these reagents was confirmed on 36 lymphoma and IS leukemia biopsy speci mens by using immunoperoxidase or immunofluorescence techniques. Additionally, flow cytometric analysis of 22 lymphoma biopsies showed that the majority of B-cell tumors were Lym-1 and/or Lym-2 positive and that within a given biopsy, a high percentage of the malignant cell population was stained. In both the immunoperoxidase and flow cyto metric studies, reactive T-cells or T-cell lymphomas were consistently negative with the exception of Hodgkin's disease tissues which, in some instances, showed a higher than expected positivity with Lym-1 and I yin-2. Approximately 40% of B-cell chronic lymphocytic leukemias were found to be positive with Lym-1 while 80% were positive with Lym-2. Immunoperoxidase staining of frozen sections of human lymphoid tissues showed that both Lym-1 and Lym-2 stained germinal center and mantle zone B-lymphocytes as well as interfollicular histiocytes. Flow cytometric analysis of normal peripheral blood demonstrated specific staining of Bcells which comprised approximately 8% of circulating lymphocytes. Immunoperoxidase staining of nonlymphoid human organs and tissues revealed weak reactivity of Lym-1 with surface colonie epithelium only. Consistent with these findings, 35 solid tumor cell lines of diverse nature were found unreactive with both Lym-1 and Lym-2. Although standard techniques have thus far failed to identify the antigen recognized by Lym-2, the membrane antigen which binds Lym-1 has been shown by iminunoprecipitation and competitive radioimmunoassay studies to be a poly morphic variant of the HLA-Dr antigen. Solid-phase radioimmunoassay techniques have shown that the antigens recognized by Lym-1 and Lym-2 are not significantly modulated after antibody exposure nor shed into the circulation of lymphoma patients. Finally, using iodine-125 labeled preparations of purified Lym-1 and Lym-2, we have determined that both reagents have a relatively large number of antibody binding sites per tumor cell and increased avidity for lymphoma cells when compared to normal and reactive lymph node B-cells. Because of the B-cell specificity of these reagents, their increased avidity for lymphoma cells, and their chemical stability after radiolabeling procedures, Lym-1 and Lym-2 appear to be promising reagents for the immunodiagnosis and therapy of the human malignant lymphomas.
An IgM-producing tumor with biochemical characteristics of a small B lymphocyte
European Journal of Immunology, 1977
A lymphocytic tumor, 38C-13, induced by the chemical carcinogen 7,12-dimethylbenz(a)anthracene in C3H/eB mice and adapted t o tissue culture, produces 7-8 S IgM with "core" carbohydrates (N-acetylglucosamines, mannoses), but not "branch" carbohydrates (neuraminic acids, fucoses, galactoses) attached t o the I.( heavy, but not to the light chains. Turnover of the 7-8 S 38C-13 IgM is slow (half disappearance time = 10-1 5 h). The IgM is released from the cells as 7-8 S IgM. The ratio of IgM synthesis t o the synthesis of all cellular glycoproteins is 0.005-0.01. After comparison of these data with data obtained with normal B lymphocytes before and after mitogenic stimulation, we conclude that 38C-13 tumor cells are transformed counterparts very near or within the population of small, mitogen-sensitive, resting B lymphocytes.
Blood, 1984
Monoclonal antibodies OKB1, OKB2, OKB4 and OKB7 have been previously shown to detect distinctive antigens displayed on B, but not on T, lymphocytes. Benign and malignant lymphoid cells were investigated for their reactivity with these antibodies in cell suspension by indirect immunofluorescence and in cryostat tissue sections by the avidin-biotin complex immunoperoxidase technique. Fetal liver pre-B cells and pre-B and common type acute lymphoblastic leukemia cells isolated from 15 patients were OKB1-OKB2+OKB4-OKB7-. All mature lymphoid tissue B cells and the neoplastic cell surface immunoglobulin-positive (SIg+) B cells isolated from each of 47 B cell neoplasms were OKB2+. OKB1 and OKB7 were expressed by interfollicular, follicular center, and many, but not all, mantle zone B cells. OKB4 was expressed by follicular center cells, but not by mantle zone or interfollicular B cells. The neoplastic SIg+ B cells isolated from 45 of 47 B cell malignancies were OKB1+OKB4+, and those isolat...