T helper cells in cytotoxic T lymphocyte development: Analysis of the cellular basis for deficient T helper cell function in the L3T4-independent T helper cell pathway (original) (raw)

T helper cells in cytotoxic T lymphocyte development: Role of L3T4+-dependent and -independent T helper cell pathways in virus-specific and alloreactive cytotoxic T lymphocyte responses

Cellular Immunology, 1990

I have compared the requirements for T helper (Th) cell function during the generation of virus-specific and alloreactive cytotoxic thymus (T)-derived lymphocyte (CTL) responses. Restimulation of vesicular stomatitis virus (XV)-immune T cells (VW memory CTLs) with VSVinfected stimulators resulted in the generation of class I-restricted, VW-specific CTLs. Progression of VSV memory CTLs (Lyt-l-2+) into VSV-specific CTLs required inductive signals derived from VSV-induced, Lyt-1'2-Th cells because: (i) cultures depleted by negative selection of Lyt-I+ T cells failed to generate CTLs; (ii) titration of VSV memory CTLs into a limiting dilution (LD) microculture system depleted of Th cells generated curves which were not consistent with a single limiting cell type; (iii) LD analysis of VSV memory CTLs did produce singlehit curves in the presence of Lyt-lf2-T cells sensitized against VSV; and (iv) monoclonal anti-L3T4 antibody completely abrogated CTL generation against VSV. Similar results were also obtained with Sendai virus (SV), a member ofthe paramyxovirus family. The notion that a class II-restricted, L3T4+ Th cell plays an obligatory role in the generation of CTLs against these viruses is also supported by the observation that purified T cell lymphoblasts (class II antigen negative) failed to function as antigen-presenting cells for CTL responses against VSV and SV. T cell lymphoblasts were efficiently lysed by class I-restricted, anti-VSV and -SV CTLs, indicating that activated T cells expressed the appropriate viral peptides for CTL recognition. Furthermore, heterogeneity in the VSV-induced Th cell population was detected by LD analysis, suggesting that at least two types of Th cells were required for the generation of an anti-VSV CTL response. VSV-induced Th cell function could not simply be replaced by exogenous IL-2 because this lymphokine induced cytotoxic cells that had the characteristics of lymphokine-activated killer (LAK) cells and not anti-viral CTLs. In contrast, CTL responses against ahogeneic determinants could not be completely blocked with antibodies against L3T4 and depletion of L3T4+ cells did not prevent the generation of alloreactive CTLs in cultures stimulated with allogeneic spleen cells or activated T cell lymphoblasts. Thus, these studies demonstrate an obligatory requirement for an L3T4-dependent Th cell pathway for CTL responses against viruses such as VSV and SV; whereas, CTL responses against allogeneic determinants can utilize an L3T4Gndependent pathway.

The role of T helper lymphocyte subsets in antiviral immunity

Research in Virology, 1993

Several lines of evidence indicate that, in mice, CD4 รท T cells can be categorized into two subclasses, Thl and Th2, based on the array of cytokines they secrete (Bottomly, 1989; Mosmann and Coffmann, 1989). In particular, activated Thl cells produce IL2 and IFNT but not IL4, ILS, IL6, whereas Th2 cells secrete IL4, IL5, IL6, but not IL2 and IFNT. Thl cells are able to induce macrophage killing of intracellular parasites, mediate delayed-type hypersensitivity (DTH) and promote IgG2a synthesis. Conversely, Th2 cells promote IgG1 and IgE production by B cells and eosinophilia.

Alterations in cytotoxic and helper T cell function after infection of T cell clones with human T cell leukemia virus, type I

Journal of Clinical Investigation, 1986

HTLV-I is a transforming human retrovirus that is an etiologic agent of adult T cell leukemia/lymphoma. To investigate the effects of this virus on T cell functions, two OKT3+, OKT4+, KI8cytotoxic clones (8.7 and 8.8) specific for allogeneic cells bearing DPw2, a class II histocompatibility antigen, were studied before and after infection with HTLV-I. The clones retained cytotoxic function for up to 70 d after exposure to HTLV-I, even without subsequent antigenic stimulation, but then lost their cytotoxic activity. Prior to infection with HTLV-I, clone 8.8 also lysed OKT3 hybridoma cells; after infection, cytotoxic activity against these OKT3-antibody bearing cells was lost in parallel with the loss of activity against DPw2-bearing target cells. In addition, expression of T3 surface antigen by HTLV-I-infected 8.8 cells was decreased at a time when they lost their cytotoxic activity, possibly contributing to the loss of cytotoxic function. Finally, clone 8.8 could provide help for nonspecific IgG production by autologous B cells when stimulated with irradiated IDPw2-bearing non-T cells. After infection with HTLV-I, this helper function became independent of DPw2-stimulation and persisted even when the cytotoxic activity was lost. An OKT4+ T cell clone thus could simultaneously manifest both cytotoxic and helper T cell activities, and these activities were differentially affected after HTLV-I infection.

Reduction of otherwise remarkably stable virus-specific cytotoxic T lymphocyte memory by heterologous viral infections

Journal of Experimental Medicine, 1996

Experimental analyses of the acute cytotoxic T lymphocyte (CTL) response to viruses have focused on studying these infections in immunologically naive hosts. In the natural environment, however, viral CTL responses occur in hosts that are already immune to other infectious agents. To address which factors contribute to the maintenance and waning of immunological memory, the following study examined the frequencies of virus-specific CTL precursor cells (pCTL) not only using the usual experimental paradigm where mice undergo acute infections with a single virus, and in mice immune to a single virus, but also in immune mice after challenge with various heterologous viruses. As determined by limiting dilution assays, the pCTL frequency (p/f) per CD8+ T cell specific for lymphocytic choriomeningitis virus (LCMV), Pichinde virus (PV), or vaccinia virus (VV) increased during the acute infections, peaking at days 7-8 with frequencies as high as 1/27-1/74. Acute viral infections such as these elicit major expansions in the CD8+ T cell number, which has been reported to undergo apoptosis and decline after most of the viral antigen has been cleared. Although the decline in the total number of virus-specific pCTL after their peak in the acute infection was substantial, for all three viruses the virus-specific p/f per CD8+ T cell decreased only two- to fourfold and remained at these high levels with little fluctuation for well over a year. The ratios of the three immunodominant peptide-specific to total LCMV-specific clones remained unchanged between days 7 and 8 of acute infection and long-term memory, suggesting that the apoptotic events did not discriminate on the basis of T cell receptor specificity, but instead nonspecifically eliminated a large proportion of the activated T cells. However, when one to five heterologous viruses (LCMV, PV, VV, murine cytomegalovirus, and vesicular stomatitis virus) were sequentially introduced into this otherwise stable memory pool, the stability of the memory pool was disrupted. With each successive infection, after the immune system had returned to homeostasis, the memory p/f specific to viruses from earlier infections declined. Reductions in memory p/f were observed in all tested immunological compartments (spleen, peripheral blood, lymph nodes, and peritoneal cavity), and on average in the spleen revealed a 3 +/- 0.4-fold decrease in p/f after one additional viral infection and an 8.4 +/- 3-fold decrease after two additional viral infections. Thus, subsequent challenges with heterologous antigens, which themselves induce memory CTL, may contribute to the waning of CTL memory pool to earlier viruses as the immune system accommodates ever-increasing numbers of new memory cells within a limited lymphoid population. This demonstrates that virus infections do not occur in immunological isolation, and that CD8+ T cell responses are continually being modulated by other infectious agents.

Mechanisms of induction of primary virus-specific cytotoxic T lymphocyte responses

European Journal of Immunology, 1992

Mechanisms of induction of primary virus-specific cytotoxic T lymphocyte responses* We have investigated the ability of various antigen-presenting cell (APC) types to induce primary anti-viral cytotoxic T lymphocyte (CTL) responses by single in vitro stimulation. Of these APC types, only dendritic cells (DC) and RMA-S lymphoma cells could induce primary CTL responses, but by divergent mechanisms. DC were capable of generating primary virus-specific CTL, either by presenting viral peptide or processed infectious virus. In contrast, RMA-S cells could not present endogenous antigen, e.g. after virus infection, but this cell line very efficiently presented exogenous viral peptides to induce primary virusspecific CTL in vitro. Spleen cells, lipopolysaccharide-induced B cell blasts or the non-mutated RMA cells did not have the ability to trigger unprimed T cells by single in vitro stimulation. We have investigated several characteristics important for primary CTL response induction by DC and RMA-S cells (summarized in Fig. 6). Primary CTL response induction by DC or RMA-S cells was blocked by anti-LFA-1 or anti-CD8 monoclonal antibodies (mAb). DC rapidly aggregated with unprimed T cells, which was independent of LFA-1 and CD8 molecules. RMA-S cells did not form conjugates with unprimed Tcells. Despite their abundant major histocompatibility complex (MHC) class I cell-surface expression, DC did not bind much exogenously added viral peptide. In contrast, the MHC class I molecules on RMA-S cells bound a large quantity of exogenously administered peptide. Powerful adhesion by DC and high expression of relevant MHC/peptide complexes on RMA-S cells are important features in the initial contact with unprimed T lymphocytes. In a later stage of contact, both DC and RMA-S cells activate LFA-1 (and CD8) molecules at the Tcell surface to strengthen and maintain the contact between T cell and APC. * This work was supported by the Netherlands Organization for Scientific Research (NWO) grants 900-500-092 and 900-507-122. Fellow of the Royal Netherlands Academy of Arts and Sciences (KNAW) .

Functional studies on the precursors of human lymphokine-activated killer cells

Cellular Immunology, 1985

Human peripheral blood lymphocytes cultured for 4 days in the interleukin 2 (IGZ)-containing cell-free supematant of the MLA144 cell line (MLA144CM) are cytolytic to NK-susceptible and NK-resistant tumor target cells. This lymphokine-activated killer (LAK) activity is dependent on IL-2 as development of LAK activity is inhibited in the presence of a monoclonal antibody (MoAb) reacting with the IL-2 receptor (anti-Tat). Addition of cyclospotin A (CyA) to mixed lymphocyte cultures inhibits the development of allospecific cytotoxic activity and inhibits the development of IL-2 responsiveness. However, development of LAK activity is unaffected by the inclusion of CyA in the cultures, showing that the LAK precursor can be functionally distinguished from the allospecific cytotoxic precursor cell. Development of LAK activity does not require mature NK cells as shown by the generation of LAK activity from NK inactive human thymocytes and lymph node cells. In addition, depletion of NK activity from human PBL does not impair the development of LAK activity. Q 1985 Academic mess, hc.

Functionally distinct human T cell clones that produce lymphokines with IL-2-like activity

Human Immunology, 1984

Various functionally distinct human T cell clones derived from in vitro mixed leukozyte cultures are found to secrete lymphokines with detectable Interleukin-2 (IL-2)-like activity upon antigenic stimulation. These lymphokine producing clones are not only dependent on exogenous growth factors provided in the form of crude phytohemagglutinin (PHA)-induced spleen conditioned medium for growth but can themselves be driven to proliferate by their own lymphokines. The induction of lymphokine production appears to be antigen-specific and the lymphokines secreted are believed to contain nonantigen and nonspecies specific IL-2-1ike activity. We show here that IL-2-like lymphokines are produced by a subset ofT lymphocyte clones, i.e., the help-independent cytotoxic T cells clones that have the capacity to proliferate to, as well as to lyse, the original sensitizing cells. In addition, other T cell clones capable of producing active lymphocytes include those clones that have the T helper cell characteristics, i.e,, can undergo antigen-induced proliferation but are not cytotoxic; in the presence of lectin (PHA), however, some helper T cells (Th) but not others, can express lectin-dependent cell-mediated lysis. Finally, yet another subset of T lymphocytes, the help-dependent cytotoxic T cell clones that cannot proliferate to antigenic stimulation, was found to produce no detectable IL-2-like activity.

Characteristics of Interleukin 2 and Interleukin 4 Dependent T Cell Lines Infected with HTLV-1 in Vitro

International Journal of Immunopathology and Pharmacology, 1997

Human T-cellieukemia/lymphoma virus type 1 (HTLV-l) is a Iymphotropic retrovirus. Cells infected with HTLV-1 in vitro, when maintained in interleukin-2 (IL-2) can be immortalized, remaining for a long time strictly dependent on IL-2 addition. In this study we have compared the effect of interleukin-4 (IL-4) and interleukin-2 on HTLV-l infection of cord blood or normal adult mononuclear cells. The results showed that either cultures of cord blood or normal adult T cells are susceptible to HTLV-1 infection in presence of IL-4 as well as IL-2. Moreover HTLV-1 infected cells in the presence of IL-4 survived only for a limited length of time in culture, while those grown in IL-2 showed the characteristics of immortalized cell lines. Moreover the profile of cytokine production showed a different pattern in HTLV-1 infected cell lines maintained in IL-4 or IL-2. This suggests that the Iymphokines differently modulate retrovirus infection in vitro.