Shared IgE-binding sites among separated components of natural rubber latex.

BACKGROUND: IgE-mediated allergy to proteins present in natural rubber latex is a well-recognized problem. Latex contains a complex mixture of proteins ranging in molecular weight from 6 to > 200 kD. OBJECTIVE: The aim of this study was to determine whether shared allergenic sites exist on separate latex components. METHODS: Following electrophoresis, latex components at 14, 24, and 46 kD were electroeluted from SDS-polyacrylamide gels and used in ELISA inhibition and immunoblot inhibition studies of human latex-specific IgE antibodies. RESULTS: A minimum of 4 major allergenic sites (for convenience labeled A through D) were found to exist in 3 components of nonammoniated latex. Minimally, 2 are present in the 14-kD component (A, B) and 3 in the 24-kD component (A-C). The 46-kD fraction has 3 or more antigenic sites (A, C, D) but lacks one (B) that is present in both the 14- and 24-kD components. CONCLUSIONS: Four different IgE-binding moieties were detected among three latex protein components (14, 24 and 46 kD). Some of these allergenic sites were shared by two or more components. Recovery of these and others from fragmented latex components will allow identification of their amino acid composition and their availability will ultimately lead to better diagnostic and therapeutic options for patients with latex allergy.

A guinea pig model of hypersensitivity to allergenic fractions of natural rubber latex.

Allergy to natural rubber latex is a growing medical problem with life-threatening aspects. The aim of this study was to learn if guinea pigs could serve as a suitable model for immediate-type hypersensitivity to latex. Guinea pigs were immunized either with whole non-ammoniated latex extract, or with one of nine SDS-PAGE-separated components. Other animals were injected with electroeluted latex components localized on gel at 14, 24 and a cluster around 45 kD. Before and after immunization, sera from the animals were examined by ELISA, immunoblots, passive cutaneous anaphylaxis (PCA) and passive systemic anaphylaxis. Latex-specific antibodies were detected by ELISA and immunoblots in sera from all immunized animals. PCA assays showed that the guinea pigs had homocytotropic antibodies dilutable to 1:250. PCA was abolished when sera from animals immunized with allergens in alum were heated at 56 degrees C for 30 min indicating the antibodies were of the E isotype. Passive systemic anaphylaxis was induced in 4 of 10 guinea pigs. The results show that guinea pigs are capable of making antibodies to latex protein components that mediate dermal and systemic anaphylaxis, paralleling the spectrum of clinical and laboratory findings of humans with immediate-type clinical latex hypersensitivity.

Regulation of chemokine gene expression by contact hypersensitivity and by oral tolerance.

In mice with hapten-induced CH, T cells of the CD4+ and CD8+ phenotypes activated the gene for JE, whereas CD8+ T cells alone caused activation of the gene for IP-10. In animals tolerized by feeding either TNCB or OX, hapten-induced expression of IP-10 but not JE mRNA was lost. The down-regulation of IP-10 gene activation was adoptively transferred from tolerized mice to naive mice by CD4+ splenic T cells. These findings reflect the differential roles of individual T-cell subsets in both enhancing and diminishing chemokine gene expression in contact hypersensitivity reactions.

Latex hypersensitivity: its prevalence among dental professionals.

Reports of hypersensitivity to latex are growing among oral health care workers, who have a high degree of exposure to latex products. The authors undertook a study to determine the prevalence of latex hypersensitivity among oral health care workers in a hospital dental practice. Among the 34 people who participated in the study, 12 percent had positive results in a skin prick test for latex. This suggests that the true prevalence rate of immediate hypersensitivity to latex in this group of oral health care workers is similar to that in other health care workers who use latex gloves frequently.

Peripheral tolerance induced in lymph nodes by syngeneic spleen cells inhibits generation of CTLs to hapten-altered self antigens but not to alloantigens.

The immunological tolerance that is induced in lymph nodes that have been exposed to syngeneic spleen cells has been examined. Development of cytotoxic T lymphocytes was used to assess the immunological status of the lymph node cells. The tolerance was studied from the viewpoint of its induction, its activation, and its specificity. We had already reported that injecting either T or B cells of splenic origin into a regional lymph node environment a week prior to immunization for CTL to hapten-altered self antigens prevents development of the CTL. Here, we confirm that syngeneic splenic cells but not lymph node cells will induce the suppression provided that spleen cells are not coupled with hapten. We now report that splenic cells that cannot replicate or synthesize and secrete protein are capable of inducing the suppression. The data suggest a preformed surface marker peculiar to spleen cells and perhaps on cells that traverse the thymus induces local tolerance that is mediated by suppressor cells. Triggering the induced suppressor T cells (previously identified as CD8-) was achieved by syngeneic spleen cells as well as by H-2-compatible, Mls-disparate spleen cells but not by syngeneic lymph node cells or apparently by allogeneic spleen cells. Furthermore, triggering suppression was achieved by hapten-coupled syngeneic spleen cells whereas such cells would not induce the suppression. Thus, activating the suppressor cells requires reexposure to splenic cells of the proper MHC haplotype, unaltered or coupled with either TNP or FITC. Once triggered, the suppression was manifested toward CTL generation against hapten-coupled syngeneic antigens on either spleen or lymph node cells but not against allogeneic antigens. Thus, the specificity of the tolerance was directed to altered self antigens despite its induction by unaltered spleen antigen. Furthermore, for suppression to be seen the spleen antigen was not required to be on the hapten-coupled syngeneic cells used for the CTL immunization. The relationship of the splenic cell "antigen" to hapten-altered self antigens and to other surface markers and its site of acquisition within the body and its significance for cell homing have become intriguing questions of importance. This information has been discussed from the viewpoint of its applicability to autoimmune diseases as well as to cessation of inflammatory reactions that may be mediated by lymph node cells.

Down-regulation of adoptive adjuvant-induced arthritis by suppressor factor(s).

We recently described a method for inducing immunologic tolerance to trinitrochlorobenzene (TNCB), a hapten that generates suppressor cells capable of down-regulating the efferent phase of TNP-specific contact hypersensitivity in rats. Peritoneal exudate cells (PEC) of such tolerized rats, upon being triggered by specific hapten, suppressed contact hypersensitivity to another hapten elicited at the same time. This implied that cells that mediate delayed-type hypersensitivity of any specificity might be down-regulated, provided that the suppressor cells are activated with specific antigen and that the unrelated delayed-type hypersensitivity is elicited in parallel. To rigorously test this possibility, we examined the ability of TNP-specific suppressor lymphoid cell factors to affect cells that mediate adoptively transferable adjuvant-induced arthritis (AIA) in rats. To induce arthritis, spleen cells from Freund's complete adjuvant-injected rats were stimulated with concanavalin A and administered to naive recipients. Prior to adoptive transfer, the cells were exposed for brief intervals to supernatants of lymphoid cells from control and hapten-tolerized rats. Supernatants of PEC and lymph node cells from hapten-tolerized rats were found to markedly reduce the effectiveness of cells that mediate AIA. The hapten-tolerized cells required reexposure to hapten prior to preparation of the supernatants. Supernatants of spleen cells from hapten-tolerized rats that had been hapten-painted as well as hapten-triggered and supernatants of lymph node cells and of PEC from only hapten-painted or hapten-triggered rats were ineffective in altering the AIA. Thus, factors from suppressor cells induced toward hapten-coupled self-antigens have been found to adversely affect the function of lymphoid cells that mediate a totally unrelated inflammatory response, namely, AIA. The clinical implications of these findings are discussed.

Induced suppression of the efferent phase of contact sensitivity in rats.

A study of the immunosuppressive systems of rats has been conducted with special attention to whether suppressor cells can be induced to down-regulate the efferent limb of contact sensitivity. Contact sensitivity (CS)1 was induced in DA rats 5 days after immunization with trinitrochlorobenzene (TNCB). Intravenous pretreatment of naive rats with TNP-coupled syngeneic spleen cells 7 days before sensitization suppressed the induction of CS by 60%. Suppression of the inductive phase of CS could be transferred adoptively into syngeneic rats with spleen cells of such tolerized animals. Cell fractionation studies showed the OX8+ (CD8) T cell population (cytotoxic/suppressor) was responsible for the suppression in the afferent phase of CS. Such cells were incapable of suppressing preexisting CS. To investigate whether suppression could be induced for the efferent phase, spleen and peritoneal exudate cells (PEC) from rats tolerized by administering TNP-spleen cells iv plus epidermal paintings with TNCB were adoptively transferred into recipients sensitized 4 days earlier. Both spleen cells and PEC suppressed the efferent phase of CS but PEC did so more efficiently. Separation of splenic cells revealed the suppressors to be CD8+ T cells. Furthermore, separation of PEC into plastic adherent and nonadherent cells showed the nonadherent (T cell enriched) cells to be noneffective alone. The adherent subpopulation conveyed suppression but did so more effectively upon addition of the T cells. Thus, T cells and macrophages may operate in concert to achieve suppression of the efferent limb of CS. PEC from tolerized rats suppressed performed CS of any specificity but only after the suppressor cells were triggered with the same antigen that induced them. Since both the afferent and efferent phases of CS have now been shown to be suppressable, two separate suppressor mechanisms may be operable in rats.

Oral administration of the contact sensitizer trinitrochlorobenzene: initial sensitization and subsequent appearance of a suppressor population.

Our earlier studies have demonstrated that intragastric administration of the hapten trinitrochlorobenzene (TNCB) 2 to 3 weeks prior to attempting sensitization with epidermally applied hapten can abrogate development of systemic contact sensitivity (CS). In this paper, we have examined whether onset of tolerance following intragastric administration of the hapten is preceded by development of hapten-specific CS. Indeed, CS was found to be present 5 days after feeding TNCB and in most experiments the response decreased significantly by Days 10 to 12. The kinetics of development of CS by the oral and epidermal routes were strikingly similar except that the magnitude of reactivity (up to 5 days) in orally sensitized mice was somewhat less than that of epidermally sensitized mice. With the exception of Peyer's patches (PP), effector cells of CS were recovered from such gut-associated lymphoid tissues as mesenteric lymph nodes (MLN), lamina propria, and lymphocytes that are present in the intraepithelial compartment of the intestinal wall. These cells as well as spleen cells of TNCB-fed mice were able to adoptively transfer CS to naive mice. The capacity of MLN and spleen cells of TNCB-fed mice to confer CS adoptively was abrogated after treating cells with anti-Thy 1.2 and anti-Lyt 1.1 antibodies plus complement thereby identifying them as T lymphocytes. Although CS decreased by 10-12 days after feeding TNCB, the decline was reversed by pretreating mice with cyclophosphamide (CY) 2 days before giving the hapten. Whereas spleen cells from animals fed hapten 5 days earlier transferred CS readily, those from mice fed hapten 12 days earlier did not. However, when 12-day spleen cells were depleted of Lyt 2+ cells their ability to adoptively transfer CS was restored. These observations indicate that feeding TNCB to mice initially produces CS, mediated by Thy 1.2+, Lyt 1.1+ lymphocytes. CS is subsequently down-regulated by activation of Lyt 2+ suppressor cells, precursors of which are sensitive to CY.

Immunosuppression in murine renal cell carcinoma. I. Characterization of extent, severity and sources.

Four cell-mediated immunological responses related to tumor elimination have been examined in mice injected with a transplantable renal cell carcinoma (Renca). Lymphokine-activated killer (LAK) cells generated in vitro from spleen cells of normal mice were capable of attacking Renca, EL-4, P815 and YAC-1 targets, but those from mice bearing Renca for 3 weeks could not. Natural killer activity, stimulated in vivo by administering poly(I) poly(C), was less than 50% of normal in Renca-bearing hosts. In addition, development of cytotoxic T lymphocytes to allogeneic targets was markedly inhibited in mice possessing the renal tumor. Finally, the delayed hypersensitivity response to a dermally applied hapten was approximately 70% less than normal in tumor-bearing mice, no matter whether the tumor existed subcutaneously or intrarenally. A kinetic study of the development of non-responsiveness using the LAK assay showed onset of poor response at 1 week, which became maximal within 3 weeks following receipt of tumor subcutaneously. The immunological depression was seen to be attributable in part to suppressor cells present among spleen cells but not bone marrow cells of tumor-bearing hosts. The suppressor cells prevented in vitro LAK generation by normal spleen cells and, when adoptively transferred to normal mice, they inhibited natural killer stimulation and delayed hypersensitivity generation. Another source of immunological down-regulation was provided by Renca cells themselves. Incorporation of Renca cells that had been X-irradiated with 30,000 rad into cultures of normal and Renca-derived splenic cells suppressed replication of both almost completely. Furthermore, the presence of X-irradiated Renca cells in cultures of normal spleen cells prevented development of LAK cells. Thus, the suppression seen in Renca-bearing mice derives from multiple sources and whether each is in any way related to the other has been discussed. Identification of the phenotypes of cells responsible for the lymphoid cell-mediated suppression and examination of its elimination are communicated in the companion paper.

Immunosuppression in murine renal cell carcinoma. II. Identification of responsible lymphoid cell phenotypes and examination of elimination of suppression.

In our companion paper we have reported that cell-mediated immunity of mice bearing renal cell carcinoma is profoundly suppressed. The non-responsiveness of such animals was found to be attributable to Renca cells themselves and to splenic lymphoid cells that down-regulate other fully capable lymphoid cells. In this communication the lymphoid cell source of suppression within Renca-bearing mice has been explored with the aim of identifying phenotypes of the responsible cells, the manner by which suppression is mediated, and initial ways by which suppression may be eliminated. A plastic-adherent cell bearing the Thy1.2 surface marker as well as the Lyt1 and Lyt2 antigens has been found to operate, perhaps in conjunction with macrophages, to down-regulate lymphokine-activated killer (LAK) cell development for natural killer (NK) and non-NK targets that include Renca cells themselves. The splenic suppressor cells lost the capacity to suppress the NK response of normal recipient mice upon shallow irradiation (250 rad) prior to adoptive transfer. Spleen cells, presumably macrophages, from Renca-bearing mice were found to suppress the generation of LAK and NK cells in vitro by synthesizing prostaglandins. Indomethacin, a prostaglandin synthetase inhibitor, blocked the induction of suppression both in vitro and in vivo, suggesting the presence of endogenous prostaglandins in Renca-bearing mice. The suppression seen in Renca-bearing mice that derives from multiple sources and has been prevented by two separate methods has been discussed from the viewpoint of the inter-relatedness of the sources.

Suppressor T cells induced by epidermally applied hapten are located in bone marrow.

Mice painted epidermally with trinitrochlorobenzene (TNCB) develop delayed type dermal hypersensitivity within five days, however, subsequent immunization with TNP-syngeneic antigens (TNP-H2k) does not generate CTLs to the same antigens. These studies were undertaken to determine what prevents development of the CTL. We report here, that a suppressor cell for CTL generation is found in the bone marrow (BM) of TNCB-painted mice. This suppressor cell is not present in spleens or lymph nodes, but is readily detected by adoptive transfer of BM cells. The cell responsible for suppression is an Lyt2+ (CD8+) T cell. Further studies with two monoclonal antibodies (one directed to a T cell suppressor factor [mAb 14-12], and the other directed to a suppressor T cell inducer factor [mAb 14-30]), demonstrated that the suppression could be reversed by either antibody when they were given prior to epidermal hapten painting. However, when each mAb was administered to recipients of BM cells from hapten painted donors, only mAb 14-12 reversed suppression of CTL generation. Examination of the number of resident BM cells revealed that TNCB-sensitized mice had 35% more cells than normal controls. When cultured in vitro with inactivated, syngeneic TNP-thymocytes, BM from normal mice readily developed TNP-self specific CTL, whereas whole BM from hapten-painted mice did not. The inability to generate CTLs was found to be attributable to suppressor cells, since BM cells from hapten-painted mice prevented CTL development by splenic T cells in culture. BM cells from normal mice did not suppress CTL generation. Suppression in vitro was not overcome by the presence of exogenous IL-2.(ABSTRACT TRUNCATED AT 250 WORDS)

Feeding trinitrochlorobenzene inhibits development of hapten-specific cytotoxic T lymphocytes by interfering with helper T-cell function.

The development of hapten-specific cytotoxic T lymphocytes (CTLs) in mice that were made tolerant by administering the hapten trinitrochlorobenzene (TNCB) intragastrically was examined. The generation of CTL specific for hapten-modified self-antigens in vivo was reduced in animals fed TNCB three times at weekly intervals prior to immunization for CTLs. In addition, splenic cells from hapten-fed mice were unable to develop CTLs in vitro. The inhibition of CTL formation was strictly trinitrophenyl-self-specific as responses to fluorescein isothiocyanate-self- or allogeneic stimulators were fully developed. Limiting dilution analyses revealed that the lack of development of CTL in hapten-fed mice was not attributable to a diminution in the frequency of precursor CTL specific for hapten altered self-antigen. By contrast, the inability of spleen cells of hapten-fed mice to produce CTLs in vitro was reversed by incorporating Lyt1+ cells from normal mice or preformed helper lymphokines into culture. That the nonresponsivity in hapten-fed mice was attributable to nonfunctioning helper cells became additionally evident when splenic L3T4+ cells from hapten-fed mice, in contrast to those from normal mice, were found to be unable to assist normal Lyt2+ splenic cells develop hapten-specific CTLs. Furthermore, fed-L3T4+ cells were ruled out to be directly suppressive of CTL production. This work illustrates that hapten-specific helper T-cell function becomes defective following orally administered hapten and that this deficit contributes to suppression of hapten-specific CTL production.

Down-regulation of cytotoxic T lymphocyte development by a minor stimulating locus-induced suppressor cascade that involves Lyt-1+ suppressor T cells, IA- macrophages, and their factors.

Down-regulation of the development of CTL has been studied in mice both in vivo and in vitro. To generate CTL to hapten-altered self Ag in vivo, an immunization protocol has been used in which the host's Th cells are stimulated by a minor locus histocompatibility Ag (Mlsd) and its precursor CTL are activated by trinitrophenylated syngeneic spleen cells. Injecting the H-2 compatible Mls-disparate spleen cells along with the TNP-coupled self cells into the hind paws causes TNP-self specific CTL to appear in popliteal lymph nodes within 5 days. We have previously reported that inducing Ts cells by i.v. injecting Mlsd-bearing cells prevents in vivo generation of TNP-self specific CTL after immunization in this way. Here the induced Ts cell as well as the mechanism by which it functions have been further examined. The suppression was seen to extend to allogeneic as well as TNP-self Ag, provided the Mlsd-tolerized animal was reexposed to Mlsd-bearing cells at the time of immunization for CTL. By transferring the Mlsd-induced suppression adoptively we have learned that the splenic suppressor cell bears Thy-1.2 as well as Lyt-1.1 Ag and inhibits the generation of CTL at the afferent limb. In addition, Mlsd-induced PEC of Mlsd-tolerized mice, but not of normal mice, mediated suppression of development of CTL in vivo. The active cells within the tolerized PEC have been identified as T cells and macrophages (M phi). Furthermore, PEC from mice tolerized to Mlsd suppressed generation of CTL directed toward TNP-self targets in vitro. T cells and M phi separated from PEC of Mlsd-tolerized mice achieved suppression best in culture when present together. In addition, Lyt-1+ splenic cells from tolerized but not normal mice cooperated to down-regulate CTL generation in vitro with peritoneal M phi from either tolerized or normal mice. Supernatants of 24- to 72-h cultures of PEC from tolerized mice were suppressive of CTL generation when incorporated at 40 to 50% of culture volume. Supernatants of T cells from tolerized PEC or spleen were suppressive in culture only when M phi from normal mice were also present. To achieve suppression dialyzed supernatants of M phi from tolerized mice could replace the M phi.(ABSTRACT TRUNCATED AT 400 WORDS)

Down-regulation of cytotoxic T lymphocyte generation by two distinct suppressor-cell systems.

Two distinct suppressor systems have been described that are capable of down-regulating in vivo generation of cytotoxic T cells directed toward haptenaltered self-antigens. One system, induced by hapten, involves three T cells that others have shown to function sequentially to suppress DTH. The initiator of this cascade is a T cell that is readily induced in spleens of mice injected intravenously with syngenic membrane-coupled hapten. This Ts, when triggered by the same syngeneic membrane-coupled hapten that induced it, elaborates a factor. The other two Ts arise in lymph nodes and spleens of mice painted epidermally with hapten. One of the two Ts in this set is readily armed by the factor of the first Ts. The factor confers its specificity and genetic restriction upon the accepting Ts. The latter, when properly triggered, makes a factor that is taken up by its companion Ts, which actually suppresses by way of a nonspecific factor. Whereas this Ts cascade is operative at the efferent limb of DTH, it mediates suppression only at the afferent phase of the CTL response. A distinctly different suppressor system is induced by minor locus (Mls) antigen. When Mlsd lymphoid cells are injected intravenously into Mlsc-possessing mice, an Lyt-1+ T-suppressor cell is generated that can be found in the spleen as well as among peritoneal exudate cells. This Ts interacts with macrophages to accomplish nonspecific suppression of the CTL response that is detectable both in vivo as well as in vitro. A Ts soluble product has been found to be effective to suppress CTL generation in vitro only when macrophages are present in culture. The macrophage that accomplishes suppression is I-A-. Although the afferent limb of the CTL response is down-regulated by this suppressor system, our in vitro culturing system is so structured as to make the helper T cell inactive. Thus, the mechanism of suppression must be oriented to the other early participants in the response, namely, precursor CTL, helper and differentiation factors, and/or the antigen-presenting cell.

A syngeneic splenic cell antigen induces a suppressor T cell in lymph nodes that controls cytotoxic T-cell and primary antibody responses.

The nonspecific suppression of immunological responses that is generated within host popliteal lymph nodes upon exposure to syngeneic normal spleen cells has been examined. The suppression, which had previously been described as being capable of preventing initiation of cytotoxic T lymphocytes (CTLs) to hapten-altered self antigens, arises within 3 to 7 days after injecting the spleen cells. Suppression was shown to be attributable to an induced T cell that was functional when transferred intravenously. Although the cell surface marker(s) on both splenic B and T cells that stimulates appearance of Ts has not yet been identified, the cells possessing the marker were not required to be viable to cause the induction. We have shown here that the Ts is fully functional when it is put in the antigenic site used for CTL immunization. The induced Ts has been identified as bearing the Lyt 2.1 cell surface marker. Furthermore, it has been shown to be insensitive to cyclophosphamide (CY), thus differentiating it from the naturally occurring Ts cell (TS0) that is known to be CY sensitive. In addition to preventing induction of CTLs toward hapten-altered self antigens, exposing popliteal lymph nodes to syngeneic spleen cells induced Ts capable of suppressing the primary IgM antibody response to sheep red blood cells. The Ts cells that suppressed the primary antibody response possessed the same Lyt cell surface markers and CY insensitivity as the Ts that mediated suppression of the CTL response. Thus, evidence that two dissimilar immunological reactions may be down-regulated by the same suppressor mechanism has been provided. Results of a kinetic study showed that the Ts prevented development of both the humoral and the cell-mediated immune responses by affecting their inductive phases. Possible targets for suppression that more than likely would have to be common to the two widely different immune responses have been indicated.

The suppressor T cell induced by syngeneic splenic cell antigen down-regulates hapten-specific cytotoxic T cells by elaborating a factor inhibitory for IL2-dependent cell replication.

An in vitro study has been made of the mechanism by which a suppressor T cell, that is induced in lymph nodes by a syngeneic splenic cell antigen, prevents generation of cytotoxic T cells specific for hapten-altered self antigens. When popliteal lymph node cells exposed in vivo to syngeneic splenic cells were immunized in vitro with heat-treated syngeneic TNP-coupled thymocytes and excess helper factors, the Ts remained inactive. In this condition the exposed popliteal lymph node cells routinely demonstrated approximately twice the CTL response developed by lymph node cells from normal mice. Nevertheless, when triggered in vitro by splenic antigen on either X-irradiated B or T cells, the exposed but not the normal lymph node cells exhibited reduced hapten-altered self-specific CTL responses. Furthermore, T cells within spleen cell-exposed popliteal lymph node cell populations when reexposed to splenic T cells made a factor that was found to be suppressive of CTL generation by normal lymph node cells in vitro. The nondialyzable T-cell suppressor factor (TsF) did not appear to act on lymph node precursor CTLs, nor on helper T cells but instead acted at the level of utilization of helper factors in the development of CTLs. In an examination of the effect of TsF on cellular replication, TsF was found to be nontoxic for CTLL-20, an IL-2-dependent T cell, and it did not hinder the uptake of IL-2 by receptor blockade of this cell. Nevertheless, the replication of CTLL-20 that is IL-2 driven was diminished in the presence of TsF. Similarly, TsF was found to be inhibitory for T-cell proliferation stimulated by mitogen but had no effect on a B myeloma cell proliferative response. Thus, TsF appears to act as an inhibitor of a T cell's capability to replicate despite the presence of the stimulus for replication, namely, IL-2.

Natural cytotoxic T cells (NCTC) that differ from natural killer (NK) and natural cytotoxic (NC) cells are present in Peyer's patches of mice.

We have examined noninduced cytotoxicity of mouse gut associated and peripheral lymphoid tissues for a wide variety of syngeneic as well as allogeneic cell lines and lymphoblasts. Lymphoid cells from Peyer's patches were found to lyse these targets in a 3-hr chromium release assay whereas lymphoid cells from intestinal mucosa, mesenteric or peripheral lymph nodes, spleen, and thymus did not. The variety of targets toward which Peyer's patch cells were cytotoxic established the latter as nonspecific and H-2 unrestricted. The cell responsible for the lytic event was identified as possessing Thy 1.2 and Ia surface antigens. This naturally cytotoxic T cell (NCTC) was found to be adherent to nylon-wool but not to plastic plates. Although both natural killer cell (NK) and non-NK targets served as targets for the NCTC, the latter were further differentiable from NK cells by lack of asialo GM1 surface marker, which is present on NK cells. In addition, NCTC remained fully functional in mice given either of the drugs cyclophosphamide or cortisone. Each of these drugs, in the doses used, markedly reduced poly(I:C)-induced NK activity. Thus, NCTC differs from NK on the basis of the spectrum of targets against which it is functional, phenotypic surface markers, insusceptibility to stimulation with poly(I:C), and insensitivity to diminution by the immunosuppressive drugs cyclophosphamide and hydrocortisone. Since NCTC is a Thy 1.2 antigen-bearing cell and is detectable in a 3-hr cytotoxic assay, it also differs from the natural cytotoxic (NC) cell. NC lacks the Thy 1.2 marker and becomes detectable only in an 18-hr cytotoxic assay. Thus, NCTC is neither an NK nor an NC cell. We have discussed the possibility that the three naturally occurring cells may be related by being dedifferentiated descendants of an antigen-specific cytotoxic T lymphocyte (CTL). Alternatively, since NCTC is confined to an anatomical site prone to ample antigenic exposure and is still identifiable as a T cell, it may be in linear transition from the CTL to the NK or NC stages.

In the murine syngeneic mixed lymphocyte reaction, one T cell subset replicates in the presence of B cells or macrophages and replication is inhibited by simultaneous presence of both stimulator cells.

The murine T-non-T cell syngeneic mixed lymphocyte reaction has been examined to determine whether B cells and macrophages stimulate the same or different subpopulations of T cells. By using experiments in which replicating T cells were suicided, we found that the two different stimulators caused replication of what appears to be the same subset(s) of T cells. Since B cells and macrophages carry the same stimulating antigens (class II plus mls or others), one would expect them to stimulate the same T cell subpopulations were it not that they have been reported to stimulate two different subpopulations in humans. When B cells and macrophages were simultaneously used as stimulators, diminished T cell replication occurred. We have found the reduced response is not attributable to exhaustion of culture nutrients or to displacement of the response peak. Other possibilities to account for this marked reduction have been discussed from the viewpoint of suppression emanating from macrophages and/or T cells.

Orally induced tolerance generates an efferently acting suppressor T cell and an acceptor T cell that together down-regulate contact sensitivity.

Intragastric administration of the hapten trinitrochlorobenzene (TNCB) suppresses development of contact sensitivity (CS) to attempted epicutaneous sensitization with TNCB. Suppression induced by feeding TNCB is hapten specific and can be transferred to normal animals with lymphoid cells from fed mice. The lymphoid cells in hapten-fed mice that cause suppression of CS have been identified as Thy-1.2-positive cells in spleen and mesenteric nodes. The suppression with Peyer's patch cells from hapten-fed mice appears to be attributable to cells bearing Thy-1.2 antigen (T cell) and to cells with surface Ig (B cell). Feeding TNCB induces an efferent-acting suppressor T cell (Ts eff), as well as an intermediary acceptor T cell (T acc) with which it interacts to block adoptive transfer of CS with immune cells. Ts eff emanating from hapten-fed mice was identified by its specificity for the hapten, insensitivity to pretreatment with cyclophosphamide (CY), ability to produce soluble suppressor factor (SSF), and requirement for T acc to be functional. The presence of T acc in hapten-fed mice, on the other hand, was confirmed by its sensitivity to treatment with CY, interaction with Ts eff or SSF, and the ability to produce nonspecific inhibitor of TDTH cells. Thus, the suppressor T cells that are induced by administering the hapten intragastrically appear to function much like the cells of the suppressor T cell cascade that are induced by giving hapten via parenteral routes.