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PubMed1976

Maternal immunocompetence. I. The graft-versus-host reactivity of lymphocytes from pregnant rats and the distribution pattern of 51Cr-labeled lymphocytes in pregnant mice.

Lymphocytes from the peripheral blood, spleen, or para-aortic lymph nodes of prrimigravida L rats carrying (L X BN) F1 (LBN) fetuses are fully capable of mounting graft-versus-host (GVH) reactions in LBN F1 recipients. The reactivity of lymphocytes from interstrain pregnant (L X BN) or intrastrain pregnant (L X L) rats, or from rats postpartum from these pregnancies, is equivalent to that of normal virgin females over a full dose-response curve, ruling out both specific and nonspecific effects of pregnancy on the intrinsic GVH competence of the maternal thymus-derived (T) lymphocyte. Attempts to block GVH reactivity with serum from pregnant rats were unsuccessful. In addition, when the distribution pattern of 51Cr-labeled syngeneic and semiallogeneic lymphocytes was studied in intact primigravida mice, there was no difference between interstrain and intrastrain pregnant mice, and there was no evidence of immunologically specific 'trapping' in the para-aortic lymph nodes draining the interstrain pregnant uterus.

Animals↗
PubMed1976

Partial tolerant state against H-2 disparate cells. No impaired specific reactivity in MLC, GVH, or antibody production.

Injection of CBA mice with H-2-compatible lymphoid cells from C3H hybrids induces a specific reduction of the mixed lymphocyte culture (MLC) response of their lymphoytes. This is not the case after injection of H-2-disparate C3H-hybrid cells, presumably because they are rapidly eliminated due to the immune response of the host. This investigation shows that CBA mice injected with CBA X C57Bl cells (H-2-disparate) at an age of 0-3 days, but not older, develop a specifically reduced MLC response after infusion of C3H X C57Bl cells as adults, indicating that they were tolerant to the C57Bl-determined antigens. However, lymphocytes from such mice showed a normal reactivity against C57Bl as assessed by MLC, graft-versus-host tests, and capacity to produce specific antibodies.

Age Factors↗
PubMed1977

Anti-parental lymphocyte reactions in neonatal F1 hybrid rats.

The subpopulation of parental-strain lymphocytes responsible for the recognition of a particular F1 hybrid strain as foreign has been shown to be subject to specific, reversible inactivation after its injection into neonatal rats of that F1 hybrid strain. Neonates born to mothers that were syngeneic with the parental-strain lymphocytes under test acquired the capacity to inactivate these lymphocytes at an earlier age than did the genotypically identical reciprocal F1 hybrids. Neonates had little capacity to inactivate completely allogeneic lymphocytes. It is inferred from the difference in behavior between reciprocal F1 hybrids that the augmented ability to inactivate anti-F1 hybrid maternal-strain lymphocytes follows exposure to such cells in utero and to antibodies with anti-F1 hybrid activity in colostrum. Specific inactivation of those marauding maternal lymphocytes with anti-fetal activity is envisaged as an important means of protection of the fetus from immunological attack by the mother. On the basis of the results presented in this and the preceding paper, it has been proposed that many of the sequelae of the transfer of immunocompetent parental-strain cells to F1 hybrid animals result not from graft anti-host activity but from an F1 hybrid anti-parental lymphocyte response that has eluded normal regulatory mechanisms. These experiments also raise the possibility that regulation of auto-immune responses may be achieved by the inactivation of lymphocytes with anti-self reactivity by other lymphocytes that respond to the recognition structure required for such reactivity.

Animals↗
PubMed1977

In vivo responses of alloreactive lymphocytes stimulated in vitro. Helper-cell activity of MLR-primed lymphocytes.

Populations of mouse lymphocytes enriched in specific alloreactive cells by priming in a mixed lymphocyte response (MLR) include cells which, when injected into congenic nude mice, enable them to make alloantibody after immunization. Helper cells for the priming H-2 alloantigens (H-2b or H-2k) were enriched relative to helper cells for the other H-2 type. Furthermore, the alloantibody responses of nude mice reconstituted with lymphocytes primed twice in vitro were virtually monospecific for the priming alloantigens. These studies suggest that lymphocytes that proliferate in MLR include lymphocytes capable of giving specific help for H-2 antigens in vivo. Nude mice reconstituted with MLR-primed lymphocytes made less antibody to bacteriophage T4 and phix than mice reconstituted with unprimed cells, and fewer mice responded. Priming of cells a second time in MLR further depleted the population of phage helper cells. Similar results were sometimes, but not always, obtained when testing reconstituted nude mice for their ability to make anti-sheep erythrocyte (SRBC) responses. These results suggest that lymphocytes primed against H-2b or H-2k alloantigens do not have specificity for antigens of T4 or phix. These alloreactive cells may also lack specificity for SRBC. However, the results do not allow a definitive conclusion to be drawn.

Animals↗
PubMed1977

Effector mechanisms in allograft rejection. III. Kinetics of killer cell activity inside the graft and in the immune system during primary and secondary allograft immune responses.

The cells recovered from sponge matrix allografts and from various lymphoid organs of recipient mice were tested for their ability to perform relevant target cell lysis in vitro. In the primary response, the peak cytotoxic activity was detected 8 days after transplantation, inside the graft, in regional lymph nodes, and in the spleen. In the secondary response, the peak cytotoxic activity in the regional lymph nodes and in the spleen took place promptly, 4 days after transplantation. The peak activity inside the graft was recorded on day 7. In the primary response, the activity inside the graft had declined. In the secondary response, the cytotoxic activity in the blood was low, and significant cytotoxic activity appeared only after the activity in the blood was high and demonstrated a biphasic profile, with one peak on day 3--4 and another activity peak on day 14. Killer cells were detected inside the graft for a far longer time in the primary (4 weeks) than in the secondary (2 weeks) response. I conclude that cellular kinetics in the secondary anti-allograft response is qualitatively but possibly not quantitatively different from the primary response.

Animals↗
PubMed1978

Evidence for two types of non-specific suppressor cells activated by the graft-versus-host reaction in mice.

In vitro experiments were carried out to investigate the immunosuppressive efect of different populations of spleen cells obtained from animal experiencing a graft-versus-host reaction (GVHR). To induce the GVHR, parental lymphoid cells were injected into adult F1 hybrid mice. GVHR-activated spleen cells (GVH-sc) taken at different times post-GVHR induction were separated into adherent and non-adherent fractions and treated with anti-theta serum plus complement. The different types of GVH-SC were added to either parental (donor-type) of F1 (host-type) normal spleen cells and cultured with sheep erythrocytes in a modified Marbrook chamber. It was found that both adherent and non-adherent 10-day GVH-SC significantly inhibited the plaque-forming cell response of F1 normal spleen cells but not that of parental normal spleen cells. Significant suppression of the parental response was observed following the addition of 15-day GVH-SC or anti-theta treated adherent and non-adherent 7-day GVH-SC. The results suggest that the non-specific immunosuppressive effect of GVH-SC is mediated by GVHR-activated macrophages and B lymphocytes found in the anti-theta treated adherent and non-adherent fractions of the GVH-SC suspensions respectively.

Animals↗
PubMed1980

Mechanisms of cell-mediated cytotoxicity in mice rejecting xenogeneic human lymphoblastoid cells.

The in vivo immunization of mice with human lymphoblastoid cell line LNH13 generates direct cell-mediated cytotoxicity by spleen cells. The lytic activity appears as early as day 3 after the intraperitoneal inoculation of 7.5 x 10(6) cells and persists at least until day 11. The killer cells do not adhere to plastic and are not retained on nylon wool columns or on Degalan beads coated with mouse Ig plus rabbit-anti-mouse Ig. The effector cells are partly inhibited by treatment with anti-Thy-1.2 serum plus complement, but this inhibition appears to be non-specific since anti-serum alone or normal serum plus complement have the same effects. Heat-aggregated IgG strongly inhibits cytotoxicity, indicating that the effector cells are Fc-positive and that such receptors are implicated in lysis. Altogether, these features strongly argue for an ADCC phenomenon. The involvement of antibodies is demonstrated by the fact that eluates (56 degrees C, 30 min) from immune cells alone induce lysis in the presence of normal spleen cells as effectors. The lytic activity of these eluates can be removed by specific adsorption on protein A coupled to Sepharose beads and on the human lymphoid target cells. Positive complementation between immune and non-immune spleen cells suggest that the arming process may occur in vitro during the assay, when antibodies are released by plasmacytes.

Animals↗
PubMed1981

Specific inhibition of anti-self reactivity following exposure of neonatal rats to lymphocytes with anti-neural activity.

Not only did newborn rats prove to be resistant to the adoptive transfer of responses against central nervous system tissues following the injection of lymphocytes from rats sensitized against syngeneic spinal cord, but such recipients were also subsequently resistant as adults to the active induction of an immune response following challenge with syngeneic cord. This resistance to active immunization against syngeneic spinal cord as adults was associated with the absence of lymphocytes with anti-neural activity and with the appearance of cells able to suppress such activity, after challenge. Interference by the lymphocytes of resistant rats with anti-neural responses was restricted to those responses directed against syngeneic neural cells.

Animals↗
PubMed1984

Auto-delayed-type hypersensitivity induced in immunodeficient mice with syngeneic modified self-antigens. II. Suppressor T cells control the autoimmune response.

The control of the autoimmune response to modified self-antigens was explored, using immunodeficient mice injected with syngeneic trinitrophenylated spleen cells (TNP-SC) as an experimental model system. X-irradiated (250 rad) A mice injected with TNP-SC and footpad-challenged 7 to 14 days later with syngeneic lymphoblasts generated a delayed-type hypersensitivity (DTH) response that was expressed by footpad swelling measured 24 h, 48 h and 72 h later. Histopathological examination showed massive inflammatory infiltration in the soft tissues of the limbs with extensive necrosis. This was not observed in X-irradiated mice that received the lymphoblast challenge only. The immunological activity was transferred from the X-irradiated TNP-SC-immunized mice to naive recipients by T cells (Lyt-1+) and not by serum, thus excluding the possibility that the inflammatory reaction is mediated by antibodies. We have previously presented evidence that the differentiation status of the lymphoblasts, and not contaminants from the incubation media, was the determinant factor eliciting the DTH response of immunodeficient mice injected with TNP-SC. Since only syngeneic lymphoblasts were able to elicit the DTH response of immunodeficient mice injected with syngeneic TNP-SC, we suggested that immunological activity was directed against self-antigens, thus expressing an autoimmune reactivity. The ability of immunodeficient mice to generate syngeneic DTH was not restricted to the TNP hapten or to inbred A-strain mice. X-irradiated BALB/c mice injected with syngeneic penicillinated spleen cells and challenged with syngeneic lymphoblasts generated a significant DTH response, in contrast to X-irradiated BALB/c mice exposed to the challenge dose only. X-irradiated A mice injected with syngeneic TNP-SC and simultaneously reconstituted with syngeneic splenocytes failed to generate a DTH response after the lymphoblast challenge, indicating that the syngeneic DTH response is controlled by normal suppressor cells. The suppressor cells were characterized as T cells carrying I-Jk, Lyt-1+, Lyt-2+ and Lyt-3+ antigenic markers. The suppressor cells abrogated the syngeneic DTH response of immunodeficient mice injected with TNP-SC, even when transferred a few days after the induction of immunological activity, but not when transferred 1 h before the lymphoblast challenge, indicating that even the established immunological activity can be restrained. Various immunological aspects of these observations and the significance of the findings in illuminating human autoimmune disorders are considered.

Animals↗
PubMed1990

Quantitation of immunoglobulins produced by donor lymphocytes in transplant patients and the possible identification of donor cell-produced antiviral antibodies by imprint immunofixation.

In the present study two patients were studied in detail to estimate the quantity of IgG of donor lymphocyte origin. Using IgG subclass quantitation and Gm haemagglutination inhibition titre, we estimated that up to 3% of IgG found after transplantation may be of donor lymphocyte origin. Analysis of viral antibody patterns by imprint immunofixation suggested polyclonal production of donor-derived antibodies.

Adult↗
PubMed1995

Engraftment and humoral immunity in SCID and RAG-2-deficient mice transplanted with human peripheral blood lymphocytes.

SCID and RAG-2 deficient mice were transplanted intraperitoneally with human peripheral blood lymphocytes (hu-PBL-SCID and hu-PBL-RAG mice). Seven days after transplantation the mice were immunized with a pneumococcal polysaccharide vaccine. Flow cytometry analysis of cells from the peritoneal cavity and the spleen after 8-10 weeks revealed that human cells had more limited engraftment in RAG than in SCID recipient mice, and that more human cells were found in the spleen than in the peritoneal cavity. Functionality of the human cells recovered from these two locations was explored by the counting of human immunoglobulin secreting cells (hu-ISC). A total of 83% of the hu-PBL-SCID mice and 29% of the hu-PBL-RAG mice had detectable hu-ISC in the peritoneal cavity and/or the spleen. The kinetic profiles of human immunoglobulins in the mouse sera during the experiment showed donor dependency. More than 90% of the hu-PBL-SCID mice had detectable levels of human IgG, IgM and IgA, while 78% had detectable levels of IgE, whereas detectable levels of IgG, IgM, IgA and IgE were measured in 37%, 64%, 68% and 23% of the hu-PBL-RAG mice, respectively. Forty-seven per cent of immunized hu-PBL-SCID mice showed a human antipneumococcal IgG level that was significantly above the background level in non-immunized mice, while none of the hu-PBL-RAG mice produced any detectable levels of human antipneumococcal IgG. In short, human PBL showed a better engraftment and a better antibody response when transplanted into SCID mice than into RAG mice.

Animals↗
PubMed1976

Alloantigens defined by cytotoxic isoantisera prepared against peripheral rabbit lymphocytes.

Seven lymphocytotoxic antisera were produced in rabbits by means of isoimmunization with purified peripheral lymphocytes. The phenotypic profile of the defined lymphocyte antigens was determined in a panel of 97 rabbits. The results indicated the existence of at least two major antigenic determinants. Further typing and cross-absorptions were performed with several of the lymphocytotoxic antisera in order to demonstrate monospecificity. The anti-3 antiserum was used to determine the distribution of the respective antigens in various rabbit organs. The highest concentration of the alloantigen was contained in the spleen, followed in descending order by the lymph nodes, lung, skin, kidney, heart and liver. Fat and red blood cells (rbc) contained no detectable antigen. A two-stage physical adherence column method was used to separate T-derived from B-derived lymphocytes and each fraction was tested for the alloantigen. Alloantigen was detected on both cell types. The fact that the anti-3 antiserum never killed more than 30% of the cells suggests that it could define a subpopulation of lymphocytes.

Absorption↗
PubMed1979

Time-dose-response studies on cellular immunity.

The time-dose responses to several murine histocompatibility antigens were studied using lymphocytes as an antigen source for immunization and the speed of rejection of skin grafts as an assay of the level of immune responsiveness. It was observed that the weaker the first set response to an antigen the higher the minimal immunizing dose and the lower the tolerizing dose; thus, there was a small "immunizing window." The speed of onset of immune responsiveness and the magnitude of change in it following first antigen contact depended on the "strength" of the antigen. When the antigen was "strong" the level of response increased rapidly following first antigen contact. When the antigen was "weak" a period of tolerance appeared following first antigen contact, which then gave way to a state of immunity; the "weaker" the antigen the longer the interval and the greater the magnitude of tolerance. Increasing the antigen dose tended to magnify and prolong the tolerance. The "weaker" the antigen the longer the interval between first antigen contact and the appearance of immunity. Once established, the level of immunity was not constant. Undulations in the levels of all responses were noted; the "weaker" the antigen the larger the undulations. Possible explanations for the undulations and implications of the time-dose-response curves in immunotherapy are discussed.

Animals↗
PubMed1977

Normal lymphocyte transfer reaction in humans and its possible mechanism.

This study illustrates the development of the skin reaction in human recipients to injected allogeneic lymphocytes from two aspects: the capacity of lymphocytes to elicit the reaction, and the ability of the recipient to react. The reaction is expressed by local skin induration and is biphasic. The first phase seems to be a hostgraft reaction; the second may require proliferation of graft cells as well as the reaction of host cells. Circulating fetal lymphocytes collected from a 12-week human embryo caused no reaction in an adult recipient; those from a 15-week embryo usually caused the first phase, with little or no second phase reaction, while those from 22- and 30-week fetuses and from a mature infant caused a normal, biphasic reaction. So, usually, did 22-week splenic lymphocytes, but fetal thymic lymphocytes usually provoked no reaction in adults. In premature infants given adult cells either both phases or the second were reduced; in normal infants the second response may be reduced but after six years of age the reaction is like that seen in adults.

Adolescent↗