Syngeneic Lymphocytes Research Articles

T lymphocyte heterogeneity in the rat: III. Autoreactive T cells are activated by B cells

Evidence has been presented to show that CD4 + autoreactive T cell lines (ATs) 2 2 Abbreviations used: APC, antigen-presenting cell; AT, autoreactive T cell lines; C, complement; KLH, keyhole limpet hemocyanin; LN, lymph nodes; mAb, monoclonal antibody; Mø, macrophages; MBP, myelin basic protein; MPL.LL, MBP-specific cell line; Mops, morphilinopropane sulfonic acid; NRS, normal rat serum; PE, phycoerythrin; RbβR, rabbit anti-rat IgG; Spc, spleen cells; Spc χ, τ-irradiated (2000 rad) spleen cells; SMLR, syngeneic mixed lymphocyte reaction. in the rat require periodic stimulation with syngeneic spleen cells for in vitro proliferation. This proliferation can be blocked by treatment of the stimulator (spleen) cells with mAb to Ia antigens. Although ATs are Ia + and can activate the allogeneic MLR, they fail to be autostimulatory. Fractionation of the spleen cells revealed that ATs can be stimulated with B cells and not by macrophages, although the latter were efficient in several accessory cell functions, including antigen presentation, lectin-dependent T cell activation and allogenic MLR response. Moreover, B cells proliferated and differentiated in response to AT cells. These data are compatible with a model in which ATs respond to hitherto undetermined B cell membrane antigen(s) in association with MHC class II antigens. These results may have important implications in understanding autoimmune responses.

Macrophages but not B cells from aged mice are defective in stimulating autoreactive T cells in vitro

In the present study the effect of aging on the capacity of Ia + cells to stimulate autoreactive T cells in the syngeneic mixed lymphocyte reaction (SMLR) was investigated. Using young CD4 + T cells as responders, it was observed that unseparated whole spleen cells from aged mice had normal stimulatory activity comparable to that of young spleen cells. Interestingly, however, when purified splenic adherant cells (SAC) enriched for macrophages or splenic B cells were used as stimulators, aged SAC but not aged B cells were found to be defective in stimulating autoreactive T cells. This defect in aged SAC was not due to decreased expression of Ia antigens since the percentage of Ia + SAC and density of Ia antigen expression was similar in both young and old mice. Also, the B cells from aged mice expressed normal levels of Ia antigens. Aged SAC, when mixed with young SAC could also actively suppress the normal SMLR. However, this suppression was not due to increased prostaglandin production but was found to be associated with interleukin-1 (IL-1) regulation, inasmuch as addition of exogenous IL-1 could completely reconstitute the defective stimulatory activity of aged SAC and also abolished the suppressor activity of the SAC. Aged mice also demonstrated an intrinsic defect in the CD4 + T cells responding in the SMLR. Together, our studies on the SMLR demonstrate an age-related defect in responder autoreactive T cells and in stimulator splenic macrophages but not in the stimulatory activity of B cells.

Defective syngeneic mixed lymphocyte response reflects incidence of diabetes in NOD mice

Defective syngeneic mixed lymphocyte response reflects incidence of diabetes in NOD mice

Aldicarb treatment inhibits the stimulatory activity of macrophages without affecting the T-cell responses in the syngeneic mixed lymphocyte reaction

Aldicarb, a carbamate pesticide used exetensively throughout the United States, has been shown in several areas to contaminate drinking water at levels exceeding 100 p.p.b. Recent studies have suggested that aldicarb at levels well below these found in drinking water may lead to alterations in mammalian health. In the present study, we investigated the possible toxic effects of aldicarb on the mammalian immune system. Specifically examined in these studies were the effects of aldicarb on syngeneic mixed lymphocyte reaction (SMLR) in which CD4 + T-helper cells (autoreactive T-cells) respond to self or syngeneic Ia molecules expressed on macrophages. The effect of aldicarb was delineated at both the responder and stimulator cell-level. When C3H mice were injected intraperitoneally with a single dose of 0.1–1000 p.p.b. of aldicarb, it was observed that there was a decrease in the stimulatory functions of macrophages, as studied by decreased capacity to stimulate normal autoreactive T-cells. Further analysis revealed that the decreased stimulatory capacity of macrophages from aldicarb-treated mice was not due to decrease in the expression of Ia antigens, since flow cytometric analysis of macrophages from aldicarb-treated mice demonstrated normal levels of Ia expression. Also, cell-mixing experiments failed to demonstrate any suppressor macrophages in addicarb treated mice. Addition of exogenous interleukin-1, however, completely reconstituted the defective stimulatory activity of macrophages from addicarb-treated mice. In contrast to these effects on macrophages, it was observed that in C3H mice treated intraperitoneally with single dose of 1–1000 p.p.b. of aldicarb, there was no evidence of alteration in the ability of autoreactive T-cells to respond to syngenic Ia molecules expressed on normal macrophages. In addition, responsiveness of T-lymphocytes obtained from aldicarb-treated mice to Ia antigens was also unaltered. These data suggested that aldicarb may selectively suppress the stimulatory activity of macrophages by inhibiting IL-1 mediated signal to the T-cells without directly affecting the T-cell functions.

Relationship between the synthesis of autoimmune antibodies and the formation of clusters of B, T and APC cells during the syngeneic mixed lymphocyte reaction in BALB/c and NZB mice: A technique for isolation of the spleen autoimmune compartment of non-immunized pathogen-free mice

Cells from the spleens of non-immunized mice were cultured in horizontal tubes, rotating very slowly around their long axis. Under these conditions, the flux speed gradient of the cell suspension near the tube walls greatly increased the chances of cells coming into contact with one another. Mixed clusters of B, T and APC cells were soon found adhering firmly to the walls of the tube; cluster formation leveled off after about 3 h. The clustered cells were easily separated from those remaining in suspension and constituted a particular cell compartment comprising a maximum of 20–30 % of the total. B cells from this compartment, cultured in complete medium for 48 h, almost exclusively produced IgM antibodies. Antibodies reacting with self antigens were so numerous in the culture medium that it is likely all IgM were self antibodies. That the clusters obtained under these conditions constituted a compartment of autoimmune cells is supported by previous work which showed that 20–30 % of spleen cells secrete IgM antibodies almost exclusively. Cluster formation as a function of age was compared in NZB mice which are used as a model of lupus erythematosus, and in BALB/c mice which never manifest self-immune pathology. The number of cells found in clusters per whole spleen increased exponentially with age in NZB mice and linearly in BALB/c mice. The production of autoimmune antibodies as a function of age also increased exponentially for NZB mice and linearly in BALB/c mice, which provides further striking support for the hypothesis that the clusters formed constitute the autoimmune comportment.

Inhibition of Autoimmune Disease in a Murine Model of Systemic Lupus Erythematosus Induced by Exposure to Syngeneic Photoinactivated Lymphocytes

MRL/l mice develop progressive, virulent autoimmune disease that has many of the features of systemic lupus erythematosus. Prophylactic treatment of MRL/l mice with syngeneic photoinactivated autoimmune splenocytes improves survival and inhibits the fulminant hyperproliferation of abnormal T cells and the production of high titer anti-DNA antibody invariably found in untreated mice. The proliferation of Thy 1+ splenic T cells was significantly decreased, and prolonged retention of the response to T-cell mitogen was found in treated mice. Treatment with unmodified cells induced a partial inhibition of disease features which did not prolong survival rates. These results suggest that phototherapy potentiates a normal immunoregulatory process which enables suppression of the development of abnormal cell populations in young MRL/l mice with relatively intact immune systems.

Synergic action of lipopolysaccharide and muramyl dipeptide in immunotherapy of DBA/2 mice with mastocytoma P815

The intratumoral administration of lipopolysaccharide (LPS) and muramyl dipeptide (MDP) in combination, but not separately, resulted in necrosis and rejection of subcutaneous P815 mastocytoma nodules in DBA/2 mice with 30 to 40% survival. Previous sensibilization of animals by LPS + MDP, treatment by indomethacin, cyclophosphamide or syngeneic lymphocytes did not augment the immunotherapeutic action of LPS + MDP combination. Reinoculation of P815 cells into cured DBA/2 mice 8 months after the disappearance of the primary tumor led to rejection of new nodules with 50% survival rate. In LPS + MDP immunotherapy of these tumors two stages may be distinguished by a thrombo-necrotic stage and that of development of immunity.

Immunostimulation circumvents diabetes in [formula omitted] mice

Diabetes susceptibility in non-obese diabetic (NOD) mice may entail faulty activation of immunoregulatory cells resulting from cytokine deficiencies. Depletion of T cells prevents disease onset in these mice. Since we had previously shown that IL-2 treatment in vivo restored the ability of NOD Lt mice to produce self-restricted suppressor T cells (Ts) in a syngeneic mixed lymphocyte reaction (SMLR), we investigated the possibility that diabetes could be circumvented by treatment with immunostimulatory agents that increase cytokine production. By 20 weeks of age, 75% of vehicle-treated NOD Lt female controls had become glycosuric, while glycosuria developed in only 17% of NOD Lt females injected with human recombinant interleukin-2 (rIL-2, 250 U twice weekly) beginning at 6 weeks of age. Treatment of mice with Poly [I:C] alone [50 μg twice weekly, an inducer of Interferon (IFN)α β ] or in conjunction with rIL-2 was even more effective, completely preventing glycosuria for 20 weeks. However, therapeutic effects required continuous administration of the immunostimulants since pancreatic insulin content declined and severity of insulitis increased following cessation of treatment. IL-2 treatment increased transcription of interleukin-1 (IL-1) mRNA in peritoneal macrophages and increased lipopolysaccharide (LPS)-stimulated IL-1 secretion in comparison to controls. In the presence of stimulators from IL-2-treated mice, T lymphocytes isolated from both controls and IL-2-treated NOD Lt mice proliferated in a SMLR and acquired Ts function. Peritoneal macrophages from Poly [I:C]-treated mice exhibited increased IFNα gene transcription and LPS-stimulated IL-1 secretion. T cells isolated from Poly [I:C]-treated mice were capable of suppressing NOD-Lt T cell responses to alloantigens in a mixed lymphocyte culture without prior activation in a SMLR. Thus, Poly [I:C] treatment may recruit a different population of regulatory cells than those elicited by treatment with IL-2. However, the mechanisms by which autoreactive T-cell clones may be regulated by these two treatments in NOD Lt mice may be synergistic. These results indicate that in addition to T-cell depletion protocols, diabetes in NOD mice can be prevented by treatment with immunostimulatory agents.

Costimulatory signal provided by a B-lymphoblastoid cell line and its Ia-negative variant.

We have analyzed the requirements of highly purified, resting murine CD4+ T lymphocytes for activation mediated by the lectin Con A and by monoclonal antibodies against the CD3 and Thy-1 molecules. Our results indicate that both the Ia-positive B-lymphoblastoid cell line M12 and its Ia-negative variant M12.C3 can provide the costimulatory activity necessary for these activation pathways. The costimulatory function is preserved upon fixation with paraformaldehyde, indicating that the costimulatory molecule(s) is (are) constitutively expressed on the cell surface. Our experiments also point to interesting differences between the M12 cell line and syngeneic Ia-positive antigen-presenting cells in generating a syngeneic mixed lymphocyte reaction. Finally, we show that the CD4+ T cell-M12.C3 cell interaction can be used to screen for interesting monoclonal antibodies that affect cell function.

Glial cells as suppressor cells: Characterization of the inhibitory function

Rat retinal glial cells (Müller cells) profoundly suppress antigen-driven activation, as well as the subsequent interleukin-2 (IL-2)-dependent expansion of autoimmune and conventional-immune syngeneic T-helper lymphocytes, through a contact-dependent mechanism. Characterization of this inhibitory function showed that some activation parameters of autoimmune T-helper lymphocytes specific to the retinal soluble antigen, responding to antigen presented on syngeneic antigen-presenting cells, were differentially affected by coculture with Müller cells. In contrast to endogenous IL-2 production, IL-2-receptor generation and proliferation, the production of interleukin-3 and of interferon-gamma were not inhibited. Inhibition of IL-2-supported proliferation of cells which had already generated the IL-2 receptor was markedly potentiated in the presence of the specific antigen, in a dose-dependent manner. The extent of inhibition was proportional to the number of Müller cells in the culture. The suppression appeared not to involve a major histocompatibility complex (MHC) Class II-related interaction and could act across allogeneic and even xenogeneic barriers. Inhibition affected normal lymphocytes, including primed T cells responding to antigen or to IL-2, unprimed spleen cells responding to the T-cell mitogen concanavalin A (Con-A) and, to a lesser extent, unprimed spleen cells responding to the B-cell mitogen bacterial lipopolysaccharide (LPS). Several permanently transformed cell lines of mouse, rat and human origin were not affected. These results may suggest participation of organ-resident cells in regulation of locally occurring immune processes.

Antiviral T cell competence and restriction specificity of mixed allogeneic (P1 + P2 → P1) irradiation chimeras

Mixed irradiation bone marrow chimeras were prepared by reconstituting lethally irradiated C57BL/10 (B10) or B10.D2 mice with T cell-depleted bone marrow cells of B10 plus B10.D2 origin. These chimeras were healthy and survived well under conventional housing conditions and after experimental laboratory infections. Of a total of 17 chimeras tested, 2 died spontaneously or from the injected virus. Twelve of fifteen chimeras mounted a measurable cytotoxic T cell response to virus. Despite approximately equal percentages of B10 and B10.D2 lymphocytes in chimeras, cytotoxic T cell responses to vaccinia virus and lymphocytic choriomeningitis virus were mediated variably by either syngeneic or allogeneic donor lymphocytes; thus the H-2 type of effector T cells frequently did not correspond to the 50:50 distribution of spleen or peripheral blood lymphocytes. Cytotoxic responses were restricted exclusively to recipient H-2 type. All mixed chimeras examined were able to mount a good IgG response to vesicular stomatitis virus. These results confirm previous data suggesting that such mixed chimeras are healthy and immunocompetent and demonstrate strict recipient-determined restriction specificity of effector T cells; they also suggest that if T help is necessary for induction of virus-specific cytotoxic T cells, it does not require host-restricted interactions between helper T cells and precursor cytotoxic T cells.

Tumor-induced alteration in macrophage accessory cell activity on autoreactive T cells.

Using a tumor-model system, differences in the accessory cell capabilities on autoreactive T cells of splenic macrophages from normal and tumor-bearing hosts (TBH) were assessed in the syngeneic mixed lymphocyte reaction. Tumor development caused a drop in autoreactivity. At 0 and 7 days of tumor growth, no drop in reactivity occurred when TBH macrophages were used as accessory cells and L3T4+ autoreactive T cells from normal mice were used as responder cells. However, by day 14, there was a 32% drop in reactivity, and by day 21 only 22% of the T cell reactivity remained when TBH macrophages were used as accessory cells. Alterations in macrophage Ia antigen during tumor growth were first investigated as the potential cause of reduced autoreactivity. Before tumor growth (day 0) 59% of the splenic macrophages were found to be Ia+. Day-7 TBH macrophages showed no difference in Ia antigen expression when compared to day 0 macrophages. However, by day 14, TBH macrophages showed a 9% decrease, and by day 21 they showed a 36% decrease in the number which were Ia+. Concomitant with the decrease in the number of Ia+ cells was a decrease in the density of Ia antigen expression on day-14 and -21 TBH macrophages. In day-14 and -21 TBH macrophages, two populations were seen that were Ia+. The first had a 10%-20% decrease in Ia antigen expression per cell while the second population had a greater than 50% drop in Ia antigen expression per cell. By titrating and mixing TBH macrophages with normal host macrophages, we assessed whether they could actively mediate suppression of autoreactive T cells. A titratable suppressive phenomenon was demonstrated using day-21 TBH macrophages. In contrast, day-7 and -14 TBH macrophages titrated with normal host macrophages had no effect on the syngeneic mixed lymphocyte reactivity. Lastly, we investigated whether the macrophage-mediated suppression was caused by increased prostaglandin secretion. Addition of indomethacin to cultures increased autoreactive T cell reactivity stimulated by normal or TBH macrophages (59% and 99% increase, respectively). Although indomethacin reduced suppression mediated by TBH macrophages, autoreactivity did not return to levels induced by untreated or indomethacin-treated cells from a normal host. Taken together, the data suggested that tumor growth modulates the function of macrophage accessory cells with autoreactive T cells in at least two ways: by decreasing Ia antigen expression and by increasing suppressor activity.

Inhibition of antiskin allograft immunity by infusions with syngeneic photoinactivated effector lymphocytes

Induction of tolerance for skin allotransplantation requires selective suppression of the host response to foreign histo-compatibility antigens. This report describes a new approach that employs pretreatment of effector cells with 8-methoxy-psoralen (8-MOP) and ultraviolet A light (UVA) to render the effector cells of graft rejection immunogenic for the syngeneic recipient. Reinfusion of photodamaged cells resulted in an immunosuppressive host response that permitted prolonged retention of histoincompatible skin grafts and specifically inhibited in vitro and in vivo responses that correlate with allograft rejection.Eight days after BALB/c mice received CBA/j skin grafts, their splenocytes served as a source of alloreactive effector cells. The splenocytes were treated with 100 ng/ml8-MOP and 1 J/cm2 UVA before reinfusion into naive BALB/c recipients. Recipient mice were tested for tolerance to alloantigens in mixed leukocyte culture (MLC), cytotoxicity (CTL), delayed type hypersensitivity assays (DTH), and challenge with a fresh CBA/j graft.Splenocytes from BALB/c recipients of photoinactivated splenocytes containing the effector cells of CBA/j alloantigen rejection proliferated poorly in MLC and generated lower cytotoxic T cell responses to CBA/j alloantigens in comparison with sensitized and naive controls. Splenocytes from these hyporesponsive mice suppressed the MLC and CTL response to alloantigen from sensitized and naive BALB/c mice. In vivo the DTH response was specifically suppressed to the relevant alloantigen in comparison with controls. Moreover, BALB/c mice treated in this fashion retained a CBA/j skin graft for up to 42 d posttransplantation without visual evidence of rejection. These results indicate that the in vivo and in vitro response to alloantigen can be attenuated by pretreating the host with photoinactivated splenocytes containing the effector cells of alloantigen rejection.

Perturbation of the idiotypic network: I. Induction with multiple alloantigen stimulation

The effect of hyperimmunization on the immune network with allostimulated syngeneic lymphocytes responding to different haplotypes was analyzed. Ten different haplotypes were used to stimulate syngeneic donor mice. Control mice were multiply immunized with incomplete Freund's adjuvant alone or with syngeneic mixed lymphocyte culture-generated lymphocytes. BALB/c mice were immunized consecutively with alloreactive blasts or allogeneically stimulated spleen cells at 10-day intervals. After a rest period of 2 months, the ratio of T helpers to T suppressors was determined by immunofluorescent staining. The functional network was probed by immunizing the mice with phosphorylcholine (PC) coupled to hemocyanin. The sera were analyzed for anti-PC antibodies and TEPC15 (T15) idiotypic expression. The results demonstrated (i) a decrease in the level of anti-PC antibody titer and T15 idiotypic expression; (ii) a decrease in the number of T helper cells and an increase in the number of T suppressor cells; (iii) a loss of PC epitope specificity; (iv) an increase of IgM antibodies expressing T15 without anti-PC specificity; and (v) an elevated level of preimmune lymphocyte proliferation and Ig secretion. These results reveal a functional network linkage in the regulation of alloreactivity and antigen response and show how repeated exposure to alloantigens can induce a perturbation of the idiotypic network controlling the response of a non-alloantigen-related BALB/c strain dominant idiotype (T15).

Site of influx of inflammatory white cells into a rejecting rat renal allograft.

In this paper we demonstrate that the vascular endothelium of a normal kidney and a syngeneic kidney graft binds only poorly syngeneic or allogeneic lymphocytes in an in vitro binding assay. On the contrary, the peritubular capillary endothelium of an allograft binds almost 7 times more lymphocytes than the peritubular capillary of a normal kidney. These findings suggest that the site of influx of inflammatory capillary white cells into the graft during rejection is the peritubular capillary endothelium.

Mixed isotype class II antigen expression. A novel class II molecule is expressed on a murine B cell lymphoma.

The structures of Ia molecules expressed by two BALB/c B cell lymphoma lines, A20-1.11 (A20) and 2PK3, were analyzed in an effort to explain the differences in antigen-presenting capacity displayed by these cells. Alloreactive T cell hybridomas specific for I-Ad and antigen-specific, I-Ad-restricted T cells responded well to A20 as the APC. The same alloreactive T cell hybridomas responded weakly or not at all to 2PK3 and the responses of the antigen-specific, I-Ad-restricted T cells were consistently lower to antigen presented by 2PK3 as compared with A20. T cells restricted to I-Ed responded equally well to either A20 or 2PK3 as APC. Additionally 2PK3, but not A20, stimulated a strong syngeneic mixed lymphocyte response. Structural analyses of the Ia antigens revealed that I-A and I-E molecules were expressed by A20, whereas an I-E and a novel I-A-like molecule were expressed by 2PK3. The novel class II molecule was affinity purified from 2PK3 cells using an mAb specific for Ad beta (MK-D6), and this molecule was subsequently shown by an RIA to react with an E alpha-specific mAb (14-4-4S) as well. Chain-specific polyclonal antisera raised against I-A and I-E alpha and beta chains indicated that the 2PK3 "I-A" alpha chain reacted in immunoblot with E alpha-specific and not A alpha-specific antisera, whereas the beta chain reacted with A beta- and not E beta-specific antisera. Peptide map and partial amino acid sequence analyses indicated that the "I-A" molecule expressed by 2PK3 represented a mixed isotype structure resulting from the pairing of Ed alpha with Ad beta. By immunofluorescence staining analysis, 2PK3 did not react with an mAb specific for Ad alpha. 2PK3 was capable of limited antigen presentation through the mixed isotype molecule to I-Ad-restricted OVA-specific T cell hybridomas, although the responses induced were low compared with presentation through I-A on A20. Previous descriptions of the expression of mixed isotype class II molecules in the mouse have resulted primarily from DNA-mediated gene transfer experiments. The results presented indicate that a mixed isotype class II molecule can be expressed naturally.

The inflammatory macrophage response to murine cytomegalovirus in genetically susceptible mice

Adherent suppressor cells have often been implicated in the depression of immunocompetence following CMV infections. We have reported that high levels of cytostatic macrophages in the peritoneal cavities of infected mice correlate with genetically-based sensitivity to CMV disease, suggesting they may modulate protective immune responses. This study investigates the properties and kinetics of such cells. Genetically-susceptible BALB/c mice infected with MCMV accumulated activated peritoneal macrophages, 7 days post-infection. These cells suppressed 3H-thymidine-incorporation and lymphokine production in syngeneic lymphocyte cultures and hence appeared to have depressed accessory cell function, although interleukin-1 production and the capacity to take up colloidal gold were enhanced. The cytostatic activity was located in a low density fraction (1.05 g/ml), which was expanded by MCMV infection. The lowest density cells had higher frequencies of infection but the proportion of cells releasing virus (less than 0.2%) was below the proportion activated, as shown by the shift in the density profile or enhanced colloidal gold uptake. A comparable accumulation of cytostatic activated peritoneal macrophages occurred in mice treated with cyclosporine A, but nude mice showed macrophage activation without cytostasis, so the role of T cells is not resolved. The spleens of infected mice maintaining high levels of virus in this organ atrophied, and the remaining cells were unable to proliferate in culture. In contrast, mice clearing the virus developed splenomegaly and restricted responsiveness, which may be governed by cytostatic cells equivalent to those in the peritoneal cavity. The spread of virus to the lymph nodes was limited and MCMV-primed cells were readily demonstrable.

Evidence for the in vivo production of interleukin-3 in lethally irradiated mice transplanted with syngeneic murine splenocytes

We wish to demonstrate new evidence for the in vivo production of interleukin-3 (IL-3). Syngeneic murine splenocytes were transplanted into lethally irradiated mice. The spleen cells in these transplant mice spontaneously produced IL-3 in cultures, and IL-3-like activity was detected in the serum. Chronological changes of the Thy-1 antigen expression on the bone marrow cells, and the number of myeloid progenitors in the bone marrow from post-transplant mice correlated with the amount of IL-3 produced in vivo. These results suggest that IL-3 may be produced by activated T cells during the syngeneic mixed lymphocyte reaction induced in vivo.

Effect of interferon-γ on the abnormality of T cell activation in NZB mice

It is known that New Zealand black (NZB) mice have a defect in the autoantigen recognition mechanism by T cells. The present study was carried out to examine whether the defect could be improved by normalizing the self Ia molecule expression on thymic reticuloepithelial cells (TRC) by using interferon-γ (IFN-γ). Syngeneic mixed lymphocyte reaction (SMLR) was used as an indicator of the mechanism of autoantigen recognition. In the reaction, T cells separated from the thymus were used as responders, and machrophages separated from the spleen were used as stimulators. When the responders were cultured with TRC, in which self Ia molecule expressions had been enhanced by IFN-γ, for activation, the low level of SMLR in NZB mice was elevated close to the normal level. It was suggested, therefore, that IFN-γ could normalize the defective autoantigen expression on TRC in NZB mice and improve the abnormal T cell activation against autoantigens.

Dendritic cells and viruses

Dendritic cells (DC) are potent at presenting viral antigens in the initiation of both primary and secondary responses. DC in the lymph nodes draining the site of infection with HSV express surface antigen and can stimulate proliferation of sensitised lymphocytes [14]. In secondary stimulation, non-responsiveness in cytotoxic T cell assays of cells from mice primed to Moloney virus was also overcome by stimulation in vitro with virus-pulsed DC [16]. Marked primary proliferative and cytotoxic T cell responses were previously found only to alloantigens and to haptens, both presented on the surface of DC (for references see [3, 4]). However, DC exposed in vitro or in vivo to influenza virus stimulated primary proliferative and cytotoxic T cell responses in normal syngeneic lymphocytes in 20-μl hanging drop cultures (Macatonia, Taylor, Knight and Askonas, in press). This provides a method for analysing primary responses to viruses in vitro without the necessity of using pre-sensitised donors. Although DC may present HIV antigens to lymphocytes the DC are also susceptible to infection with HIV [15, 22]. This occurs in vivo as evidenced by the infection of Langerhans cells in AIDS patients [18]. This infection of DC may not only compromise their function in antigen presentation but also act as a reservoir of virus which is handed on to T cells during the close clustering of the presenting cells with the T cells during the initiation of the immune response. DC may thus act not only as antigen-presenting cells in the initiation of immune responses but may also be a route of immunoregulation delivering either positive or negative signals to lymphocytes.