Characterization studies of suppressor cells in murine bone marrow chimaeras.

When BALB/c bone marrow cells were transferred to lethally X-irradiated C3H/He mice either intrasplenically (i.s.) or intravenously (i.v.), suppressor cells were detected by means of MLR coculture assays in the spleen of i.s. and i.v. chimaeras. Some but not all of the suppressor cells expressed a Thy 1.2 antigen, indicating that suppressor cells either sensitive or insensitive to anti-Thy 1.2 antibody plus complement treatment were generated in the spleen of i.s. and i.v. chimaeras. According to the examination of Lyt alloantigen expression on suppressor T cells, Lyt 1+2-, Lyt 1-2+, and Lyt 1+2+ suppressor T cells appeared to be present. The culture supernatants from several T-cell clones showed the suppressor activity against alloreactive MLR. Cell surface markers of these clones were composed of Lyt 1+2-, Lyt 1+2+ and Lyt 1-2+. In addition, since Carrageenan treatment abrogated the suppressor activity of plastic dish adherent cells, we conclude that some of them were composed mainly of macrophages.     

Surface Lyt phenotype of suppressor cells in C57BL/6 mice infected with Mycobacterium lepraemurium.

C57BL/6 were infected intravenously with 10(7) Mycobacterium lepraemurium (MLM). At increasing time intervals after infection different isolated splenic cell subpopulations were tested for their ability to suppress the mixed lymphocyte reaction (MLR) of normal syngeneic mouse splenocytes. During the first 6 months after infection neither T depleted nor plastic adherent spleen cells from infected mice exerted a suppressive activity on the normal mouse allogeneic proliferative response. Conversely, splenic T cells from MLM infected mice exhibited suppressive activity as early as 2 months after infection. Attempts to characterize the Lyt phenotype of splenic suppressor T cells from 6 months infected mice showed that both Lyt 1+ 2- and Lyt 2+ enriched cell subsets possessed the ability to suppress the MLR of the normal mouse spleen cells and Lyt 1+ 2- T cells were shown to be more efficient suppressors than Lyt 2+ cells.     

The functional association of Lyt antigens with lymphokine production.

The effects of anti-Lyt antibodies on secretion of lymphotoxin, macrophage-activating factor and interferon were studied. Anti-Lyt-1 antibodies added to primary mixed lymphocyte reactions (MLR) significantly enhanced lymphokine secretion, whereas anti-Lyt-2 antibodies had no effect in primary responses. However, when added to secondary MLR, anti-Lyt-1 antibodies manifested moderate enhancing effect on lymphokine levels, whereas anti-Lyt-2 antibodies markedly reduced lymphokine secretion. These results suggested that T cells of different subsets produced lymphokines during primary and secondary responses. The Lyt phenotype of T cells involved in lymphokine secretion and the effects of anti-Lyt antibodies on selected T-cell subsets were therefore studied. It was demonstrated that the majority of lymphokine producing cells during primary MLR reside within the Lyt-1+2- population, which was potentiated by anti-Lyt-1 antibodies, but unaffected by anti-Lyt-2 antibodies. However, during secondary MLR, a considerable proportion of lymphokines was produced by the Lyt-1-2+ population, which was blocked by anti-Lyt-2 antibodies. Lymphokine production induced by concanavalin A, in contrast to that induced by alloantigens, was unaffected by anti-Lyt-2 antibodies. The implication of these results on the role of Lyt molecules in T-cell functions is discussed.     

Positive selection of T-cell subsets. I. Proliferative responses of Lyt 2 separated thymocytes and splenic T cells.

Flow microfluorometry analysis of peanut lectin non-agglutinable (PNA-) thymocytes (ThC) reveals the existence of 30%-50% Lyt 1,2,3+ and 50%-70% Lyt 1+,2,3- subpopulations. Using positive selection on anti-immunoglobulin-coated (Mage) plates, we selected PNA- Lyt 2+ and PNA- Lyt 2- ThC as well as their peripheral counterparts in the spleen. These populations were tested in parallel for their ability to respond to concanavalin A (Con A) and phytohaemagglutinin (PHA), to respond to allogeneic stimulation in the mixed lymphocyte reaction (MLR); ThC subpopulations were also tested for their ability to provide synergy with lymph node cells (LNC) in the MLR. It was found that (a) Lyt 2- cells of both thymic and splenic origin responded to all doses of Con A or PHA; (b) PNA- Lyt 2+ ThC were unresponsive to Con A or PHA, whereas splenic Lyt 2+ T cells responded to low doses of mitogens; and (c) PNA- ThC of both Lyt phenotypes responded in a MLR and provided synergy with LNC in the MLR. These data support the notion that Lyt 2+ cells of either PNA- or PNA+ subpopulations must undergo post-thymic maturation before becoming responsive to low doses of T-cell mitogens.     

Immunoregulatory abnormalities induced by experimental reovirus infection: functional alterations in T-cell subpopulations

A primary anti-sheep red blood cell (SRBC) plaque assay system was used to analyze the effect of reovirus infection on immunoregulatory T-cells. A decrease in the plaque-forming cell (PFC) response of splenic lymphocytes was observed within 24 hr of infection with 10(9) or 10(11) particles of reovirus type 1 or reovirus type 3 and persisted for more than 7 days. In coculture experiments, T-cells from infected mice were found to produce less help to control B-cells and to suppress helper function mediated by control T-cells. This suppression did not require the presence of an Lyt1,2,3 cell. In addition, isolated Lyt1 cells from reovirus-inoculated mice provided less help than did Lyt1 cells from normal controls. These abnormalities were more marked following inoculation with reovirus type 3 than with type 1. Live virus was not required for these effects, as T-cells from mice inoculated with ultraviolet (uv)-inactivated reovirus when added to normal B-cells reproduced the effects of infection with live virus. Furthermore, these changes were not the result of alterations in the percentages or numbers of distinct Lyt-bearing T-cell subpopulations in the spleens of inoculated mice. Thus, humoral hyporesponsiveness following reovirus infection of adult mice is associated both with active T-cell suppression of B-cell help and with decreased help mediated by the Lyt1 subset.     

T lymphocytes mediate protection against Yersinia enterocolitica in mice: characterization of murine T-cell clones specific for Y. enterocolitica.

Yersinia enterocolitica is enteropathogenic for humans and rodents, causing intestinal and extraintestinal diseases. The cellular immune response of the infected host has not yet been analyzed in detail. Therefore, we used a parenteral mouse infection model to determine the role of T lymphocytes in immunity against Y. enterocolitica. We report the generation and characterization of Y. enterocolitica-specific T-cell clones isolated from spleens of intravenously infected C57BL/6 mice. The T-cell clones obtained showed the phenotype of helper T cells (L3T4) or cytotoxic T cells (Lyt2). All T-cell clones were positive for the interleukin-2 (IL-2) receptor (Tac antigen, p55 subunit) and were negative for the gamma delta T-cell receptor. L3T4+ clones produced small quantities of IL-2 (less than 1 U/ml) when stimulated with heat-killed Y. enterocolitica, whereas Lyt2+ clones produced no or extremely low levels of IL-2. In contrast to IL-2 production, both L3T4+ and Lyt2+ T-cell clones produced considerable quantities of gamma interferon (500 U/ml). When transferred into nonimmune mice, some of the L3T4+, as well as the Lyt2+, T-cell clones could mediate at least partial protection against a challenge of a lethal dose of Y. enterocolitica. These data demonstrate for the first time the generation and characterization of Y. enterocolitica-specific T-cell clones and provide evidence that T cells may be involved in protection against enteropathogenic Y. enterocolitica.     

Do L3T4+ T cells act as effector cells in protection against influenza virus infection

This study aimed to analyse the roles of Lyt 2+ and L3T4+ memory T-cell subpopulations in murine influenza infection. Previous work has shown that Lyt 2+ cytotoxic T-cell (Tc) clones can adoptively transfer protection. We therefore wished to see whether L3T4+ (Th) cells could also act as protective effector cells. Donors for adoptive cell transfer were thymectomized mice, depleted in vivo of either Lyt 2+ or L3T4+ T cells with monoclonal antibodies (MAb) and then infected with influenza virus (A/X31). Primed spleen cells, after removal of the B cells, were transferred into irradiated hosts infected simultaneously or persistently with a heterologous influenza virus and the effect on lung virus replication determined. Depletion of L3T4+ T cells suppressed the formation of IgG antibodies after influenza virus infection, indicating significant depletion of T-helper function. Yet Lyt 2+ class I MHC-restricted Tc cells were effectively primed in these mice, albeit to half the normal level. Adoptive transfer of the Lyt 2+ memory T cells cleared virus in a persistent infection within 6 days. Spleen cells selected for L3T4+ T cells cleared virus within 21 days of transfer in a simultaneous infection and reduced viral titres in a persistent infection, but not as effectively as L3T4+-depleted spleen cells. Although no Lyt 2+ cells were detected by fluorescence staining in Lyt 2+-depleted spleens, we could detect low levels of class I MHC-restricted influenza-specific Tc memory cells in host spleens following influenza infection. Therefore, whether the early viral clearance is solely due to L3T4+ T cells is not clear. Lyt 2+ memory T cells appear more efficient in this respect than L3T4+ memory T cells.     

Effects of first-order Cryptococcus-specific T-suppressor cells on induction of cells responsible for delayed-type hypersensitivity.

Cell-mediated immunity is an important aspect of host resistance against Cryptococcus neoformans. Using a CBA/J murine model, we demonstrated that injection of cryptococcal antigen (CneF) at dosages sufficient to stimulate the antigenemia observed in cryptococcosis patients induces specific T-cell-mediated suppression of the cryptococcal delayed-type hypersensitivity response. The purpose of this study was to establish whether Lyt 1+, first-order T-suppressor (Ts1) cells block the induction of T cells responsible for delayed-type hypersensitivity (TDH cells) or whether they function by inducing Lyt 2+, efferent suppressor (Ts2) cells. In one set of experiments, suppression was observed when Ts1 cells were adoptively transferred to recipient animals the day before, the day of, or the day after immunization; however, when Ts1 cells were transferred after TDH cells were present, no suppression occurred. In other experiments, putative TDH cells from lymph nodes (LN) or spleens were adoptively transferred from mice after immunization or after a suppressive dose of CneF or adoptive transfer of Ts1 cells and immunization. Delayed-type hypersensitivity could not be transferred with LN or spleen cells from mice receiving the suppressive dose of CneF or the Ts1 cells, even when the LN or spleen cells were treated with anti-Lyt 2.1 antibody and complement to remove any Ts2 cells. Delayed-type hypersensitivity was readily transferred with LN or spleen cells from immunized mice whether the cells were or were not treated with anti-Lyt 2 and complement. Furthermore, the cells in the tolerized LN cell pools responsible for suppression of TDH cell induction were Lyt 1+ 2-, I-J+ cells, which is the phenotype of the Ts1 cells. Taken together, these data indicate that Ts1 cells inhibit the induction of TDH cells. This finding, coupled with the previous demonstration that Ts1 cells or a Ts1 cell-derived soluble factor (TsF1) induces Ts2 cells, establishes that the cryptococcal Ts1 cells are bifunctional in the suppressive pathway.     

Immunoregulatory Pathways in Adult Responder Mice

This report describes the alteration of helper-suppressor balances in an immune response (Ir) gene-controlled system by varying the route and form of antigen injection. Adult responder BALB/c mice develop Lyt 1+2-, T cells for delayed-type hypersensitivity (DTH), and T-cell proliferative (Tprlf) responses to subcutaneous injection of either poly(Glu60Ala30Tyr10) (GAT)-coupled syngeneic spleen cells (GAT-SP) or GAT emulsified in complete Freund's adjuvant. In contrast, intravenous injection of adult responders with GAT-SP results in specific unresponsiveness for DTH, Tprlf, interleukin-2, and plaque-forming cell (PFC) responses. This tolerance is mediated by both suppressor T cells (Ts) and a functional clonal inhibition. Lyt 1-2+ Ts suppress the induction (afferent limb) of GAT-specific DTH and PFC but not Tprlf responses. The reduced T-cell proliferation observed in GAT-tolerant mice is due to a non-transferable mechanism(s), possibly functional clonal inhibition. Our data are compatible with a multi-step pathway involving both proliferating and non-proliferating helper T (Th) cells. In addition, the fine specificity of tolerance induction for DTH and Tprlf responses was examined by using the related antigens poly(Glu60Ala40) (GA) and poly(Glu50Tyr50) (GT). Tolerance is exquisitely specific, as GA tolerizes responses to GA and GAT, whereas GT tolerizes GAT but not GA responses. Thus, both the route and form of antigen administration are important to the induction and regulation of immune response in Ir gene-controlled systems. Possible mechanisms governing the Th/Ts balance and the induction of GAT-specific tolerance and suppression for cellular and humoral responses in adult responders are discussed.     

T-cell-mediated immune response in murine malaria: differential effects of antigen-specific Lyt T-cell subsets in recovery from Plasmodium yoelii infection in normal and T-cell-deficient mice.

For analysis of the role of immune T cells in protective immunity against murine malaria, Plasmodium yoelii-immune Lyt T-cell subsets were functionally characterized in vitro and in vivo. Selected Lyt2- and Lyt2+ T cells from P. yoelii-immune C57BL/10 mice differed in their capability to proliferate in response to P. yoelii antigen in vitro. Only the Lyt2- T-cell population produced T-cell growth factor upon restimulation, and none of the selected T-cell subsets produced detectable amounts of macrophage activating factor. Lyt2- but not Lyt2+ lymphocytes were capable of transferring protection to normal C57BL/10 mice. When transferred into T-cell-deficient C57BL/6-nu/nu mice, adoptive resistance to P. yoelii by Lyt2- lymphocytes was only demonstrable after prior reconstitution of recipients with normal T cells. These results suggest an interaction between P. yoelii-immune Lyt2- T cells and normal T lymphocytes via T-cell growth factor in the development of protective immunity to malaria.     

Immunological Consequences from Exposure to Benzo(a)pyrene During Pregnancy

Progeny and maternal immune status after benzo(a)pyrene (BP) exposure of mothers at midpregnancy is disrupted in fetal liver (FL), in spleen and in thymus during pregnancy and postnatally. Mice suffer deficiencies in splenic and thymic mixed lymphocyte responses (MLR), and disorientations of T antigen expressing cells, punctuated by exorbitant increases in Lyt2, especially in FL, FL Lyt2 do not suppress an MLR, while Lytl mediate suppression. Isolated Thyl show a weak response to Concanavalin A: FL Thyl weakly express an MLR. Maternal macrophages and progeny B cells are also functionally abnormal. Thus, BP Induces generalized immune deficiency that may affect ontogeny and which is potentially deleterious to health.     

Heterogeneity of cytolytic and suppressor clones of alloactivated cells generated from soft agar colonies

Long-term cell cultures (or clones) were developed from soft agar colonies of lymphocytes alloactivated in mixed leukocyte culture reaction (MLR). Two types of colonies were identified: upper colonies that grew on the agar surface, and lower colonies found within the agar layer. Virtually all cytolytic clones originated exclusively from the upper colonies. Two groups of cytolytic clones could be distinguished, one with strong and the other with weak proliferation upon restimulation. Upper clones were capable of inhibiting primary MLR proliferation and this appeared to be related to their cytolytic effect on the stimulator. Many noncytolytic lower clones were found to suppress primary MLR cultures. Considerable heterogeneity was apparent from differences in the magnitude of suppression and the ability of the clones themselves to undergo stimulator-induced proliferation. Kinetic studies of MLR suppression were conducted to further analyze this heterogeneity. Two major kinetic patterns were observed. One showed a biphasic proliferation pattern of the MLR + clone culture. The first peak appeared to reflect an enhanced proliferation of the clone. The second phase seemed to represent diminished proliferation of the MLR responder. This type of suppression may be related to T cell growth factor depletion from MLR by the proliferating clone. The other kinetic pattern showed a consistently low proliferation of the MLR + clone culture throughout the 8-day assay period. Subsequent testing of these suppressor clones in third-party MLR cultures suggested that the specificity of suppression was unrelated to HLA-DR, MB, MT, and SB.     

Production of macrophage-activating and migration-inhibition factors in vitro by serologically selected and cloned Listeria monocytogenes-specific T cells of the Lyt 1+2- phenotype

Lyt-selected Listeria-immune T lymphocytes from peritoneal exudates and cloned T cells were cocultured with heat-killed listeriae and peritoneal macrophages from nonimmune donors. Supernatants were assayed for: activation of macrophages for tumoristatic and tumoricidal activity via macrophage-activating factors and migration-inhibition factor activity. Peptone-induced peritoneal macrophages were activated by incubation with the supernatants for 24 h. For examination of cytocidal activity, 51Cr-labeled EL4 tumor cells were subsequently added, and 51Cr release was determined. Cytostatic activity was measured by adding unlabeled EL4 tumor cells to the pretreated macrophages and determining [3H]thymidine incorporation 24 h later. Migration-inhibition factor production was examined in an agar microdroplet assay. Only Listeria-specific T cells of the phenotype Lyt 1+2- proved active in these assays, whereas T cells of the phenotype Lyt 1-2+ were not active. When T-cell clones were used, a single clone was capable of inducing macrophage-activating and migration-inhibition factor production at cell concentrations of ca. 10(3)/ml.     

Antigen-specific suppression of human antibody responses by allogeneic T cells. II. Cell interactions involved in the generation of suppression

Specific antibody responses to influenza virus were obtained in vitro from human blood mononuclear cells (PBMC). Antibody production in these cultures was profoundly suppressed by the addition of allogeneic T cells with the surface phenotype Leu2a+ (CD8+), Leu8-. Suppression by allogeneic T suppressor (Ts) cells required interactions only between T-depleted B (E-) cells and allogeneic Leu2a+. No evidence was obtained for T-T cell interactions, or for Ts inducer cells similar to those described for nonspecific antibody responses to pokeweed mitogen. Moreover, allogeneic E+, or allogeneic Leu2a+ cells were able to suppress specific antibody responses by E- cells when help was provided by T cell-replacing factor showing that the target of suppression was the responding E- cells, and not T helper cells. In contrast to allogeneic T cells, allogeneic E- cells did not suppress antibody production when added to cultures of unfractionated PBMC (E- + E+). That is, Ts cells activated to allogeneic E- were unable to suppress antibody production by the syngeneic E- cells present in the same culture tube. This result shows that alloactivated Ts cells were specific for the allogeneic E- target cells, and that suppression was not mediated by nonspecific allogeneic effects. Allogeneic Ts cells therefore differ from Ts cells in pokeweed mitogen responses by their specificity, and by their activation in the absence of Ts inducer cells.     

Complexity of effector mechanisms in cyclosporine-induced syngeneic graft-versus-host disease.

Administration of the immunosuppressive drug cyclosporine after syngeneic or autologous bone marrow transplantation elicits a T-lymphocyte-dependent autoimmune syndrome similar to graft-versus-host disease (GVHD). The onset of this autoaggression syndrome, termed syngeneic GVHD, is associated with the development of a highly restricted repertoire of CD8+ autoreactive T cells that recognize a peptide from the invariant chain, termed CLIP, presented by major histocompatibility complex (MHC) class II molecules. Clonal analysis reveals 2 distinct subsets of autoreactive T cells defined by their activation requirement for either the N-terminal or the C-terminal flanking regions of CLIP and by their cytokine profile. The studies here reveal that the autoreactive T-cell clones requiring the N-terminal flanking region of CLIP produce type 1 cytokines (interferon [IFN]-gamma, interleukin [IL]-2, and tumor necrosis factor-alpha). In contrast, the autoreactive T-cell clones that require the C-terminal flanking region of CLIP produce type 2 cytokines (IL-4, IL-10, transforming growth factor-beta). As assessed in a local graft-versus-host reaction assay, the N-terminal flanking-restricted clones mediate changes consistent with acute GVHD, whereas the clones responsive to the C-terminal flanking region do not. Moreover, the autoreactive T-cell clones restricted by the C-terminal flanking region of CLIP ameliorate the pathogenic potential of the cells responsive to the N-terminal flanking region of CLIP. The mechanism accounting for this regulatory affect appears to be the downregulation of mRNA message for type 1 cytokines (IFN-gamma and IL-2). The C-terminal-restricted autoreactive T-cell clones, however, could manifest disease with dermal changes similar to those seen in chronic syngeneic GVHD, provided that IFN-gamma was present. Consistent with these observations was the demonstration that type 1 cytokines are preferentially detected during the acute phase of syngeneic GVHD, whereas type 2 cytokines dominate during the chronic phase. The results suggest that acute and chronic syngeneic GVHD is mediated by distinct autoreactive T cells, which are separated by their fine specificity for the CLIP-MHC class II complex and by their cytokine profiles.     

The rat mixed lymphocyte reaction: roles of a dendritic cell in intestinal lymph and T-cell subsets defined by monoclonal antibodies.

Cells present in the intestinal lymph of rats were obtained in large numbers by removing the mesenteric, portal and caecal lymph nodes and cannulating the thoracic duct 6 weeks later. About 1% of the cells present in the thoracic duct lymph of these mesenteric lymphadenectomized rats had striking dendritic morphology, were strongly Ia+ but labelled weakly with monoclonal antibodies that recognize rat B or T cells. It was found that intestinal lymph was highly enriched for cells that stimulated allogeneic T cells in the mixed lymphocyte reaction (MLR) and cells with stimulator activity co-purified with dendritic cells. Thus, these dendritic cells appear phenotypically and functionally similar to the dendritic cells that have been described in the mouse spleen and rat lymph node. The ability of the intestinal lymph cells to stimulate rat T cells was used to determine which of the two subsets of these cells were the prime responders in the rat MLR. These subsets, defined by monoclonal antibodies, have been shown by previous work to display close functional analogies to the Lyt 2+ and Lyt 2- subsets in the mouse and to the two human T-cell subsets that have been defined by monoclonal antibodies. It was found that the T-cell subset that contains the helper cells for antibody responses proliferated when irradiated, fully allogeneic or semi-allogeneic thoracic duct cells were used as stimulators, but the subset containing suppressor T cells did so only in the fully allogeneic system. Detailed studies showed that in the absence of helper cells in the responder population T cells in the stimulator population of helper phenotype were responsible for proliferation of the suppressor T-cell subset observed in fully allogeneic MLRs. Proliferation of the suppressor T-cell subset could be obtained using semi-allogeneic stimulators, provided that the F1 cells were derived from a source containing dendritic cells but it was shown that, as in the case with fully allogeneic stimulators, the helper T cells in the stimulator population were playing an active role. These results demonstrate that proliferation of the suppressor T-cell subset in the rat MLR is dependent on blastogenic activity provided by the helper T-cell subset and suggest that in some situations this blastogenic activity may arise through the recognition, by the helper cells, of environmental antigens presented on dendritic cells. It has been reported that in the human MLR both T-cell subsets proliferate but that only the helper subset does so when antigen-primed cells are stimulated with specific antigen. The present experiments, by emphasizing the activity of helper T cells in the stimulator population in the MLR, cast doubt on the implication that recognition of alloantigens in vitro differs in an essential way from that of soluble antigens.     

Type-1 and type-2 CD8+ T-cell subsets isolated from chronic adult periodontitis tissue differ in surface phenotype and biological functions.

Cloning of CD8+ T cells expressing the alpha beta T-cell receptor from inflamed human gingiva revealed that at least two different subsets were found within the tissue and that these subsets were able to interact with each other. One subset produced high levels of interferon-gamma (IFN-gamma) and no interleukin-4 (IL-4) or IL-5, exhibited phytohaemagglutinin (PHA)- or anti-CD3-mediated cytolytic activity, and were CD28+. The other subset produced high levels of IL-4 in combination with IL-5, displayed no cytotoxicity and were CD28-. From the latter subset CD8+ T-cell clones were able to suppress the proliferative response of cytotoxic CD8+ T-cell clones. This suppression could be abolished by anti-IL-4 monoclonal antibodies. However, IL-4 alone was not able to induce the suppression. Our results indicate that CD8+ T cells might participate in local immune responses by the suppression of IFN-gamma-producing cells and by favouring humoral responses via the production of IL-4 and IL-5.     

Inside the thymus, Mls antigen is exclusively presented by B lymphocytes.

The ability to stimulate an Mls-1 mixed lymphocyte reaction (MLR) is predominantly expressed by low density B lymphocytes in the spleen and peritoneal cavity of normal adult mice, and is absent in splenic B cells 1 month after lethal irradiation and reconstitution from autologous bone marrow. Coreconstitution of these mice with normal syngeneic peritoneal cells restores the stimulatory potential of splenic B cells, but sorted CD5+ or CD5- IgM+ lymphocytes from peritoneum are equally good stimulators, suggesting that functional Mls-1 expression may require long life spans and selection. Bone-marrow-reconstituted DBA/2 mice that fail to express Mls-1 antigens in the periphery nevertheless maintain T-cell receptor V beta 6 and 8.1 deletions among the newly formed T cells. These findings led us to directly investigate the Mls stimulatory ability of purified antigen-presenting cell populations inside the thymus. We report here that thymic B lymphocytes seem to represent the only intrathymic cell population able to stimulate Mls-1 MLR.     

The role of antigen presentation in the ontogeny of immune responses to herpes simplex virus.

The ability of epidermal antigen presenting cells (APC) to induce immune responses to herpes simplex virus (HSV) has been studied in mice of differing ages. Using an in vivo model of HSV infection we have demonstrated that neonatal epidermal cells (EC) induce specific suppression of DH to HSV in normal syngeneic adult mice. The suppression is transferable and mediated by T cells of the Lyt1+, Lyt2-, L3T4+ phenotype. The ability of EC to induce suppression persists up to 4 weeks of age, whereas EC from mice 6 weeks of age or older induce positive DH responses to the virus. This correlates with the susceptibility of mice of the different ages to HSV infection and their ability to mount DH responses to the virus.     

Mouse T cell clones against Mycobacterium avium: identification of clones that modify resistance against atypical mycobacteria infection.

Mouse T cell clones against live Mycobacterium avium were generated from the spleens of BALB/c mice infected with M. avium TMC 702. Eighth clones were of the L3T4+ subset, whereas two were of Lyt2+ subset. Six of the L3T4+ T cell clones were of the TH1 subset whereas two were of the TH2 subset, judged on the profile of cytokine release. One of the Lyt2+ clones exhibited significant cytotoxicity against M. avium-infected mouse macrophages. Transfer of clones to nude BALB/c mice infected with M. avium was associated with insignificant changes in resistance for seven clones. One clone, of the L3T4+/TH2 subset, transferred significant resistance to the infection, also associated with infusion of supernatants from the clone, which was fully inhibited by neutralizing with anti-interleukin 4. By contrast, infusion of one TH1 clone and the cytolytic Lyt2+ led to increased microbial growth in the spleens and livers of infected mice, which was not apparent on infusion with supernatants. Application of clones' supernatants on infected macrophages had marginal effects on M. avium growth and was not correlated with protective or suppressive activity. Overall, these results suggest that T cells may influence M. avium growth in vivo in a bidirectional manner and also suggest that interleukin 4 may be an important factor in host resistance to M. avium.