Interactions between primed and unprimed cells in the regulation of in vitro antibody responses. I. Role of “plasma cells” as inducers of suppression

Specific immune suppression has been shown to be activated in culture by the interaction of primed and unprimed T cell subsets. The primed cell involved is found 8 days after immunization in spleen but not in lymph node or thymus cell populations. When the primed spleen cells were fractionated by nylon wool passage or anti-Thy-1 plus complement (C) treatment, prior to culture with unseparated unprimed cells, suppression was detectable only with primed B cells present in the co-cultures. Treatment of the primed spleen cells with anti-PC. 1 (an antiserum specific for plasma cells) plus C eliminated their ability to cooperate with either unseparated or T cell-enriched populations of unprimed cells in suppressing the antibody response of the co-cultures. These data are consistent with the hypothesis that antibody-secreting plasma cells activate suppressor T cell precursors in cell populations not previously exposed to antigen.     

Rapid loss of feedback suppressor T-cell activity after priming in vivo.

T cells from antigen-primed mice have a diminished capacity to mediate feedback suppression when compared to T cells from unprimed mice. This was demonstrated using an in vitro model of B-cell induced feedback suppression in which spleen cells from mice primed with sheep erythrocytes (SRBC) activate feedback suppressor T-cell precursors to mediate suppression of the primed spleen cell response. The addition of splenic T cells from unprimed mice to cultures of spleen cells from SRBC-primed mice resulted in suppression of the secondary IgM and IgG anti-SRBC response. In contrast, no suppression was detected when T cells from mice primed with SRBC were added to the primed spleen cell cultures. The loss of suppression by T cells from primed mice was antigen-specific and was detectable by 24 hr after priming, coinciding with the appearance after priming of T-cell enhancing activity. The reduced suppressive activity could be due to changes in the active T-cell subset itself or to the appearance of cells or factors within the T-cell population that block or mask detection of feedback suppression. In either case, the present finding suggests that priming of a host not only activates feedback suppression induction mechanisms, but also rapidly affects the ability of the T-cell population to develop effective feedback suppression.     

The suppressive effect of carrier priming on the response to a hapten-carrier conjugate

Carrier-primed cells, having some of the properties of T lymphocytes, have been found to inhibit the primary anti-hapten response of spleen cells to challenge with a hapten carrier conjugate. The antibody response of CBA/Ig^b spleen cells in irradiated CBA (Ig^a) recipients was measured by means of a radioimmunoassay using ¹²⁵I-labeled anti-Ig^b antibody. It was found that the primary anti-2,4,6-trinitrophenyl(TNP) response to TNP-KLH (keyhole limpet hemocyanin) was suppressed in recipient mice primed with KLH as compared with similarly treated unprimed or bovine serum albumin-primed recipients. This suppression was transferred by injection of 10⁸ spleen cells from CBA mice primed with KLH seven days beforehand, but not by injection of serum from such mice. The suppressive effect was abolished by treating the carrier-primed spleen cells with anti-T cell sera and complement before transfer. Priming with KLH seven days beforehand had only a small suppressive effect on the response to an unrelated antigen, DNP-ovalbumin, but there was marked suppression if KLH was again administered with the unrelated antigen. It is considered that the suppressive effect is specific in induction, but nonspecific in expression and that it is a manifestation of a homeostatic mechanism limiting the extent of the immune response.     

Adoptive suppression of erythrocyte autoantibodies in Lyt-2+ depleted recipients

Cells suppressing red blood cell (RBC) autoantibody responses were found to exert their effect in Lyt-2+ depleted recipients. Mice injected with monoclonal antibodies to Lyt-2 became deficient in Lyt-2+ cells as judged by indirect immunofluorescence. These mice and normal recipients were given either rat RBC primed spleen cells (suppressor cells) or normal spleen cells and challenged with rat RBC. The autoantibody response of both Lyt-2+ depleted and normal recipients was suppressed as compared with that of mice given normal spleen cells. It is suggested that an Lyt-1+ cell may be involved in the expression of suppression.     

Allotype suppression induced in the adoptive transfer system: the variables of the system and an apparent absence of a role for T cells.

A study has been made of the variables concerned in allotype suppression of adult spleen cells in the adoptive transfer system. These are; SRBC (antigen) dose; the dose and timing of injection of anti-allotype serum IgG; the number of spleen cells transferred and whether these cells were taken from primed or unprimed donors. Adoptively transferred primed cells are considerably less susceptible to suppression by concomitantly injected anti-allotype serum IgG than are unprimed spleen cells. Injection of anti-allotype serum during the period after adoptive transfer, has shown that primed cells loose their susceptibility sooner (2 days) than the unprimed cells (4 days). Allotype heterozygous CBA spleen cells are less susceptible heterozygous CBA spleen cells are less susceptible to allotype suppression than either allotypically homozygous or heterozygous non-H-2k cells (H-2b,d, or s). Allotype suppression of the TI IgG response to DNP-Ficoll was measured 7 days after adoptive transfer of allotype-homozygous cells from both normal and nude CBA mice (unprimed). The results indicate that T cells do not play a role in the initiation of short-term allotype suppression in the adoptive transfer system.     

Induction of suppressor cell activity by cyclosporin A and/or uremic serum in normal versus uremic peripheral blood mononuclear cells

We investigated induction of suppressor cell activity in peripheral blood mononuclear cells (PBMC) from normal or uremic subjects. The cells were primed with cyclosporin A (CS-A), uremic serum, or both and subsequently cocultured with fresh phytohemagglutinin-stimulated normal allogeneic responders. CS-A-primed normal as well as uremic PBMC significantly suppressed responder cell thymidine incorporation. Uremic serum primed normal, but not the uremic, PBMC exerted a significant suppression on responder cell proliferation. No significant additive suppressive effect was detected following coculture of responders with PBMC primed by CS-A and uremic serum together. Exclusion of a distinct suppressor cell subset by panning, using anti-Leu-2b monoclonal antibody, did not result in blunting suppressive activity of the CS-A or uremic serum pretreated cells. On the other hand, adherent cell depletion of primed PBMC completely abolished their subsequent suppressive effect on responder cell proliferation.     

Immunosuppression caused by antigen feeding. I. Evidence for the activation of a feedback suppressor pathway in the spleens of antigen-fed mice

Sheep red blood cells (SRBC) administered by the oral route to normal mice elicited no detectable splenic anti-SRBC plaque-forming cell (PFC) response until 8 weeks of antigen feeding. At this time a splenic IgA anti-SRBC PFC response was detected. On the other hand, spleen cells taken from mice given oral SRBC for 1-5 weeks showed striking changes in their in vitro anti-SRBC responsiveness as compared to spleen cells from normal mice. This was evidenced by enhanced early (days 3-4) in vitro responses, followed by suppressed late (day 5-6) in vitro responses. Both early enhancement and late suppression were T cell-mediated. Early enhancement appeared to be mediated by helper T cells of the Lyt-1+2.3- phenotype. Late suppression was also mediated by Lyt-1+2.3- cells, but Lyt-2-bearing cells had to be present in culture for suppression to occur. Lyt-2-bearing cells could be replaced with normal T cells. Furthermore, elimination of cells bearing I-J-encoded determinants from the T cell population isolated from the spleens of antigen-fed mice also partially relieved suppression. Thus, antigen feeding appears to activate a feedback suppressor pathway in which Lyt-1+2.3-, I-J subregion determinant-bearing T cells can suppress immune responses by causing normal T cells to become suppressor effectors. No evidence was found to show that antigen feeding induced Lyt-1-2.3+ suppressor cells in the spleen, nor were any serum suppressor factors detected.     

Characterization of suppressor-inducer cells which control the production of rat erythrocyte-induced anti-erythrocyte autoantibodies

Mice immunized with rat erythrocytes produce autoantibodies to their own red blood cells, distinct anti-rat agglutinins and autoantigen-specific suppressor cells. Suppressor cells were detected by adoptive transfer of rat erythrocyte-immunized spleen cells to naive recipients. Such recipients failed to make erythrocyte autoantibodies after immunization with rat erythrocytes although their anti-rat erythrocyte response was unimpaired. Depletion and enrichment studies were performed to identify the cell type(s) which transfer suppression. B cell depletion of rat erythrocyte-immunized spleen cells by passage over Ig/anti-Ig-coated bead columns abrogated the transfer of suppression. However, suppression was still transferred after rat erythrocyte immunized spleen cells were passed over beads coated with a complex of 4-azido-2-nitrophenyl (NAP)-mouse IgG-rabbit IgG anti-NAP suggesting that T cells bearing Fc gamma receptors are not responsible for suppression. Positively selected B cells from rat erythrocyte-immunized spleen cells caused some suppression of erythrocyte autoantibodies but only after high numbers of cells were transferred. Neither positively selected Lyt-1+2- nor Lyt-1-2+ T cell subpopulations transferred suppression. By contrast, rat erythrocyte-immunized spleen cells which contained a mixture of B memory and T cells were suppressive and retained their suppressor activity after removal of Lyt-1-2+ but not Lyt-1+2- cells. It is proposed that these Lyt-1+2-T cells belong to a distinct population of suppressor-inducer cells which together with memory B cells stimulate the generation of effector T suppressor cells in naive recipients.     

Selective participation of immunoglobulin V region and major histocompatibility complex products in antigen binding by T cells.

Antigen-binding inhibition studies using microscopic autoradiography were performed on T or B cell-enriched lymphocyte populations. Antibodies specific for the "framework" of immunoglobulin heavy or light chain variable domains (VH or VL), or anti-H-2 and anti-Ia antisera were used. T cell subclasses were separated with anti-Lyt antisera and complement. It was found that antigen-binding T cells of different subclasses can be inhibited selectively with only one of the two anti-V region antibodies. Antigen binding to Lyt-1+ cells was inhibited by anti-VH, while Lyt-2+,3+ cells were inhibited by anti-VL specifically. Anti-Ia antisera inhibited unprimed Lyt-1+ antigen-binding cells, whereas anti-H-2K or anti-H-2D anti-sera inhibited unprimed Lyt-2+,3+ antigen-binding cells, and both classes of immune T antigen-binding cells.     

Mice naturally tolerant to C5 have T cells that suppress the response to this antigen

We examined whether C5-sufficient mice which are naturally tolerant to this antigen have suppressor T cells to C5 humoral immune response. Two congenic strains of mice B10.D2 (NSN) and B10.D2 (OSN) differing only in the presence or absence of C5 were used. Irradiated (760 rds) sufficient hosts were reconstituted with a nonadherent spleen cell suspension from either sufficient or deficient mice or a mixture of both. Hemolytic C5 levels were assayed. Sufficient spleen cells appeared to prevent the drop of C5 level caused by anti-C5 antibody made by deficient spleen cells. Spleen cell suspensions from sufficient mice primed with deficient spleen cells exhibited better anti-C5 activity than normal sufficient spleen cell suspensions. This anti-C5 activity is abrogated by treatment of the NSN spleen cell suspensions obtained from NSN primed with OSN spleen cells with anti-Thy-1.2 antiserum and complement. Suppression of the humoral response to C5 failed to affect the anti-sheep red blood cell immune response. Suppressor T cells are resistant to low-dose irradiation, cortisone treatment and adult thymectomy. In contrast, they are sensitive to high doses of irradiation and both high and low doses of cyclophosphamide treatment. Thus, C5-sufficient mice, in contrast to C5-deficient mice, appear to have antigen-specific suppressor T cells which downregulate the humoral immune response to C5. In addition, we examined the relationship of these suppressor T cells to the state of tolerance in helper T cells of C5-sufficient mice. This was done in irradiated deficient mice which were repopulated with spleen cell suspensions selectively depleted of either Lyt-1+ or Lyt-2+ T cell subsets. These chimeras were challenged with murine C5 and both the primary and secondary immune response was measured by inhibition of the C5 hemolytic activity. It was found that only spleen cell suspensions of the deficient mice selectively depleted from the Lyt-2+ subset of T cells responded to the antigen both in the primary and secondary response. In contrast, either subset of T cells from the sufficient mice failed to respond. Thus, it appears that in sufficient mice helper T cells to C5 are intrinsically tolerant or physically and/or functionally deleted. In conclusion, the data suggest that both T cell compartments are unresponsive and play a role in the mechanism of tolerance to a physiologic antigen.     

New immunomodulator lobenzarit disodium (CCA): activation of IgE class-specific suppressor T lymphocytes and regulation of IgE antibody response enhanced by sublethal X-irradiation in SJL/J mice

The effect of the new immunomodulator lobenzarit disodium (CCA, disodium-2,2'-iminodibenzoate) on IgE antibody response was studied in X-irradiated SJL/J mice. IgE antibody response to dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH) was enhanced by sublethal X-irradiation (400 R), and this enhanced IgE production was suppressed by unprimed normal spleen cell transfer. Oral administration of CCA, every day from 1 day after immunization with DNP-KLH to 1 day before bleeding out, reduced IgE antibody response in sublethally X-irradiated SJL/J mice. The suppressive effect of CCA in the mice was exerted at an early stage of the IgE antibody response. Moreover, CCA showed IgE class-specific suppression and did not suppress IgG class antibody production. CCA-induced reduction of IgE antibody response in SJL/J mice seems to be mediated by suppressor T cells, since depletion of T cells by treatment with antithymocyte serum and complement abolished the CCA-induced reduction and splenic T cells from CCA-treated mice were able to transfer the suppressive effect.     

In vitro functional characterization of SAG-3--a naturally occurring monoclonal antibody in NZB mice: specificity toward functional T cell subsets

Natural thymocytotoxic autoantibodies (NTA) have been suggested to contribute to the T cell mediated immune dysfunction prevalent in NZB mice. However, direct studies to confirm these suggestions have been difficult to perform due to the absence of large amounts of pure NTA. To overcome this problem, we generated a naturally occurring monoclonal antibody from an NZB mouse which possesses the serological characteristics of serum-NTA. Herein, we report the in vitro functional characteristics of this monoclonal antibody designated SAG-3. In vitro treatment of spleen and lymph node cells from BALB/c mice with SAG-3 and complement eliminated most of the proliferative response to Con-A and moderately reduced the response of these cells to PHA-P and PWM. Complement mediated cytolysis of SAG-3 reactive cells significantly reduced the capacity of unprimed lymph node cells to generate primary CTL responses in vitro against allogeneic cells. However, only a partial decrease in CTL activity was observed when primed secondary CTL effector cells were treated, due primarily to the depletion of CTL's specific towards MHC class I antigens. Furthermore, a moderate reduction in antigen-specific helper T cell function was observed. Pretreatment of responder cells with SAG-3 + C' did not appear to reduce the proliferative MLR response to a wide range of stimulator cells. In addition, contrary to previous reports on serum-NTA, SAG-3 appears not to be reactive against Con-A induced suppressor T cells. Finally, by FACS analysis SAG-3+ thymocytes constitute an overlapping population of Lyt-2+ and L3T4+ thymocytes. These findings are discussed in comparison to previous reports on NTA.     

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.     

In vitro activation of natural killer-like cytotoxicity by specifically in vivo primed T-helper lymphocytes in the rat.

The interaction between the rat non-cytotoxic T lymphocyte subset, which is marked by the W3/25 monoclonal antibody, and natural killer cells was investigated. Specifically in vivo primed lymph node cells were restimulated in vitro with the priming antigen and co-cultured with a source of natural killer cells and their precursors. Cytotoxic activity, generated during a 4 day incubation period, was assessed by lysis of a rat natural killer cell-sensitive tumour target cell line, y3Ag123. This cytotoxic activity was more fully described as natural killer cell cytotoxicity on the basis of target cell specificity, using a range of natural killer cell-susceptible and -resistant targets. The W3/25-positive T cell, separated from the in vivo primed lymph node cells by nylon wool column elution, antibody labelling and sorting on the fluorescence-activated cell sorter, was shown to be necessary to stimulate the generation of this activity. W3/25-negative cells were not active in this respect. The activation was shown to be mediated via lymphokine(s), probably interleukin-2, present in concanavalin A-induced lymphocyte culture supernatants. These supernatants could be used to substitute for in vivo primed, restimulated W3/25-positive lymph node cells in activating natural killer cell cytotoxicity from normal spleen cells. Nylon wool column-eluted spleen cells, activated in vitro with conditioned medium were separated into OX8-positive and OX8-negative subsets using the fluorescence-activated cell sorter. The distribution of cytotoxic activity related to that of freshly derived rat natural killer cells.     

Characterization of a third-order suppressor T cell (Ts3) induced by cryptococcal antigen(s).

Previous studies from our laboratory have shown that a high dose of cryptococcal culture filtrate antigen (CneF) administered intravenously induces a complex suppressor cell cascade which down-regulates the cell-mediated immune response to Cryptococcus neoformans antigens. The primary objective of this investigation was to determine whether a suppressor cell induced by immunization is required for efferent suppression of the cryptococcal delayed-type hypersensitivity (DTH) response. Our approach to this problem was to immunize CBA/J mice with CneF emulsified in complete Freund adjuvant and then 6 days later to collect spleen cells from the immunized mice and adoptively transfer these cells along with C. neoformans-specific second-order suppressor T cells (Ts2) to naive syngeneic recipients at the time of footpad challenge of the recipients with CneF. To establish which populations of cells in the spleens of immunized mice play a suppressive role, mass cytolysis with specific antibodies and complement was performed before the spleen cells were transferred to naive animals. Since the phenotype of the cells responsible for the transfer of the cryptococcal DTH response had not been completely determined, we first demonstrated that the cells responsible for DTH were L3T4+ Lyt-2- cells. Subsequently, we established that a Thy-1+ L3T4- Lyt-2+ I-J+ cell population induced by immunization was required along with C. neoformans-specific Ts2 cells for efferent suppression of the cryptococcal DTH response. In addition, we demonstrated that the suppressor cells in the immune cell population were derived from cyclophosphamide-sensitive precursors. These data indicate that a third suppressor cell population is required for efferent suppression of the cryptococcal DTH response. As in the azobenzenearsonate and 4-hydroxy-3-nitrophenyl acetyl hapten suppressor models, the Ts2 cells in the circuit mediate their effects through this third suppressor component. Since the mode of induction and the phenotype of the third C. neoformans-specific suppressor cells are similar to those reported for Ts3 cells in other antigen-specific suppression models, we referred to this third suppressor cell in the C. neoformans-specific suppressor cell cascade as a Ts3 cell.     

Leukocyte subpopulations which amplify or suppress antigen-induced proliferation in Syrian hamsters

The proliferative response of spleen cells, obtained from Syrian hamsters sensitized to hen egg albumin emulsified in complete Freund's adjuvant, is lower in magnitude than the response of draining lymph node cells. In this study, the cellular regulatory mechanisms which may lead to splenic hyporesponsiveness were examined. Although unfractionated spleen cells were not suppressive, the addition of nylon wool nonadherent normal spleen cells to sensitized draining lymph node (target) cells markedly suppressed antigen- but not mitogen-induced proliferation. Suppressor cell activity was not detected in normal lymph nodes. Suppression could be overcome by culturing splenic suppressor plus target cell mixtures in the presence of large quantities of antigen. Suppressor cell activity was radioresistant. In addition to nonadherent suppressor cells, the hamster spleen also contains an adherent cell population(s) which amplified antigen-induced proliferation. Adherent cells with amplifying activity were also present in lymph nodes. The addition of adherent cells abrogated splenic suppression of proliferation. Collectively, these data indicate that the hamster spleen contains both suppressive and amplifying leukocyte subpopulations which may be involved in the regulation of the immune response to certain antigenic stimuli.     

MHC- and Igh-unrestricted interaction between L3T4+, Lyt-2- Ts inducer cells and L3T4-, Lyt-2+ Ts effector cells is required for T cell-dependent suppression of DTH to histocompatibility antigens

This paper describes the characteristics of T suppressor inducer (Ts ind) cells which can interact with T suppressor effector (Ts eff) cells and thereby can account for suppression of delayed-type hypersensitivity (DTH) to alloantigens. Adoptive transfer of spleen cells from mice intravenously (i.v.) injected with allogeneic spleen cells one day earlier induced an antigen-specific state of suppression in the recipients. This became apparent when DTH was induced by subcutaneous (s.c.) immunization of the recipients three days after transfer. The induction of suppression after adoptive transfer of spleen cells required Thy-1+, L3T4+, Lyt-1+2- cells. These cells that by themselves did not exert a suppressive effect induced a state of suppression in recipient mice by activation of recipient-type Ts eff cells. Therefore, the former cell type was classified as Ts ind cell. When athymic nude mice were used as recipients, Lyt-2+ precursors of Ts eff cells had to be transferred together with the Ts ind cells to induce a state of suppression in these mice. The Ts ind cells could activate Ts eff cells in MHC- and Igh-incompatible recipients. The results are discussed in relation to previously described immunoregulatory T cell pathways.     

The immunosuppressive effect of monoclonal anti-Lyt-1.1 antibodies in vivo

Monoclonal anti-Lyt-1.1 alloantibody was produced as tissue culture supernatant and administered to mice. The antibody, given intraperitoneally, resulted in the suppression of all T cell functions studied, but was without direct effect on B cells. Thus, skin and tumour allograft survival was prolonged and there was suppression of the delayed-type hypersensitivity response; T cell help inthe anti-sheep red blood cell antibody response, responder cells in the mixed lymphocyte reaction (MLR), leucoagglutinin-responsive cells, cytotoxic T cell (Tc) function and the induction of Tc were either totally or partially suppressed, all these responses being mediated by Lyt-1+2- or Lyt-1+2+ cells in CBA/H mice. By contrast, there was no inhibitory effect on the MLR-stimulating or lipopolysaccharide-responsive cells. The administration of the anti-Lyt-1.1 antibody was accompanied by a depletion of Lyt-1.1+ T cells from both spleen and lymph node. These studies indicate that the monoclonal anti-Lyt-1.1 antibody is active in vivo with a selective effect on T cells. The results also have important implications for studies of T cell interactions in the mouse in vivo, and for similar studies in man.     

Antigen-specific helper T cells are essential for cytotoxic T cell responses to metabolically inactivated stimulator cells.

Primed spleen cells respond well to metabolically inactivated stimulator cells while normal spleen cells do not. This observation has been interpreted as showing that cytotoxic T cell precursors are different from unprimed precursors in their antigen recognition requirements for induction. A different model is proposed here which accounts for these observations as due to enhanced helper cell levels in primed populations. Experiments are described in this study which test several predictions of this model. These experiments show that in the presence of in vitro primed helper T cells, normal cells are able to respond efficiently to glutaraldehyde-fixed stimulator cells. The helper effect is antigen-specific. Since unprimed spleen cells can be efficiently induced by metabolically active stimulators (γ-irradiated cells) and can respond to glutaraldehyde-fixed antigen (metabolically inactive cells) only in the presence of specific helper cells, it seems reasonable to propose that helper cell signals are enhanced by a nonantigenic property of γ-irradiated stimulator cells requiring metabolic activity. It is also clear that glutaraldehyde-fixed cells are anti- genically intact as helper cells, primed to antigens on γ-irradiated stimulator cells, efficiently and specifically help a response to fixed stimulators. Conversely, helper cells primed in vitro to glutaraldehyde-fixed stimulators recognize antigen on γ-irradiated stimulator cells. The level of help generated in response to glutaraldehyde- fixed stimulator cells is at least 10-fold higher in primed cells than in normal cells. In addition, primed spleen cells can be induced in vitro to yield helper function by both fixed or unfixed stimulator cells. Normal helper cell precursors are induced at least 100-fold more efficiently by γ-irradiated as compared to glutaraldehyde-fixed stimulator cells. This work supports the idea that a major effect of priming, which allows primed cells to respond to metabolically inactive stimulators, is to enhance levels of helper T cells in the primed population.     

T cells bearing unique membrane antigen and their regulatory role in humoral immune response in mice

The mutant lpr gene causes a generalized massive T cell proliferation and an associated autoimmune disease in mice. The lpr T cells bear unusual membrane phenotypes and play a causative role in the pathogenesis of autoimmune disease through excess production of some lymphokines. Therefore, the "aberrant" T cells in lpr mice may be normal regulatory T cells in non-lpr-bearing mice, if nonproliferative. We thus attempted to produce a xenogeneic monoclonal antibody (ALP-1) directed against the lpr T cells. The ALP-1 monoclonal antibody reacted with the antigen Lp-1 distributing on about one half of the proliferating lymph node cells from MRL/Mp-lpr/lpr mice but not on the cells from non-lpr-bearing mice. Nevertheless, a significant number of Lp-1+ cells appeared in the lymph node and spleen cells from these normal mice after in vitro stimulation with mitogens. Functional studies revealed that either the treatment with ALP-1 plus complement of the antigen-primed splenic T cells from non-lpr-bearing mice or the addition of ALP-1 alone all but completely abolished the development of plaque-forming cells against sheep red blood cells in T-B cell coculture. Pretreatment of spleen cells with ALP-1 plus complement, however, did not affect other in vitro T cell functions such as mitogen-induced proliferative responses, interleukin 2 production, response in mixed lymphocyte culture (MLC) and the MLC-induced cytotoxic T cell activities. Thus, Lp-1+ T cells in normal mice are unique T cells that may play an important regulatory role in humoral immune response.