Experimental models for prevention of graft-versus-host reaction in bone marrow transfusion. II. Inability to prevent graft-versus-host reaction in an H-2 identical combination.

Splenomegaly was strong in the degree and continued for a long period of time in adult F1 hybrids between AKR (H-2k) and C3H/He (H-2k) mice after transfer of spleen cells from normal C3H/He mice. In spleen cells of such F1 recipients, cytotoxicity was detected by an in vivo neutralization test using methylcholanthrene-induced sarcoma or AKR origin as target cells. All of newborn F1 recipients died within 17 days after cell transfer. Induction of splenomegaly and cytotoxicity was not prevented by repeated pretreatments of donors with sonicated AKR spleen cells in saline, which suppressed completely such phenomena of graft-versus-host reaction in an H-2 nonidentical combination. Induction of cytotoxicity in the spleen of F1 recipients was not prevented by a pretreatment of donors with AKR spleen cells in complete Freund's adjuvant, which suppressed the induction of cytotoxicity in an H-2 nonidentical combination. Graft-versus-host reaction appears to be stronger in a combination between parental strains of which major histocompatibility antigens were identical.     

Genetic regulation of delayed-type hypersensitivity responses to poly (Tyr,Glu)-poly(DLAla)--poly(Lys): expression of the genetic defect in the induction and manifestation phases in H-2s and H-2f mice.

The genetic defect of H-2s and H-2s non-responder mouse strains in both the induction and manifestation phases of delayed-type hypersensitivity (DTH) responses to poly(LTyr,LGlu)-poly(DLAla)--poly(LLys)[(T,G)-A--L] was analysed. Utilizing an in vitro system to activate DTH effector T cells, we observed that non-adherent T cells of (H-2f X H-2b) F1 or (H-2s X H-2b)F1 responder mice, could not be activated on antigen bearing adherent cells of H-2f or H-2s haplotypes. On the other hand, these T cells were effectively sensitized on adherent cells derived from either F1 or parental (H-2b) responder mice. These results indicate that in these mouse strains the genetic defect, in the induction phase of DTH, is expressed at the level of the antigen presenting cell. In subsequent experiments, we were able to "correct' the non-responsiveness of H-2s recipients by transfer of educated and irradiated (H-2s X H-2b)F1 T cells together with normal F1 adherent cells. Normal non-adherent and nylon wool enriched T cells failed to restore these responses. Similarly, antigen-pulsed F1 irradiated peritoneal exudate cells could stimulate DTH responses in SJL recipients of (SJL X C57BL/6)F1 (T,G)-A--L educated cells. The genetic defect of H-2s mice in the manifestation phase of the DTH reaction is thus also expressed on the antigen presenting cell.     

Host Defenses in Experimental Scrub Typhus: Genetics of Natural Resistance to Infection

Genetic resistance to lethal infection with Rickettsia tsutsugamushi was studied in over 30 inbred strains, inbred hybrids, and outbred stocks of mice. Inbred mice infected intraperitoneally with the Gilliam strain of R. tsutsugamushi showed three patterns of response: susceptible (A/HeJ, C3H/HeDub, C3H/HeJ, C3H/HeN, C3H/St, CBA/J, DBA/1J, DBA/2J, and SJL/J), resistant (AKR/J, BALB/cDub, BALB/cJ, C57BL/6J, C57L/J, and SWR/J), and selectively resistant (A/J). The selectively resistant pattern was characterized by random deaths occurring throughout the titration range and was also observed in three of the six outbred mouse stocks surveyed. No correlation was evident between the H-2 haplotype of inbred mice and their response to Gilliam infection. The progeny from five different Gilliam-resistant by Gilliam-susceptible inbred parental crosses were all resistant. Study of F(1), F(2), and parental backcross generations of BALB/cDub (resistant) and C3H/HeDub (susceptible) hybrids indicated resistance was dominant and was controlled by a single gene or a closely linked cluster of genes that were autosomal and not linked to coat color. The resistance of BALB/cDub mice was not due to an inability of host cells to support rickettsial growth, since C3H/HeDub and BALB/cDub embryo cell cultures supported similar growth of Gilliam organisms. C3H/HeDub mice, although susceptible to intraperitoneal Gilliam infection, were capable of mounting an immune response to Gilliam antigens, since subcutaneous infection was not lethal and did protect animals against subsequent intraperitoneal challenge with either the Gilliam or Karp strains of R. tsutsugamushi.     

Immunobiological studies on experimental visceral leishmaniasis. II. Adherent cell-mediated down-regulation of delayed-type hypersensitivity response and up-regulation of B cell activation.

Visceral leishmaniasis or kala-azar is characterized by a variety of immunopathological consequences in man. The most remarkable of these are the depression of cell-mediated immunity and polyclonal B cell activation. The consequences observed in man could be induced in a murine model by inoculating the causative agent, Leishmania donovani. The cell-mediated response was studied in this murine model in terms of the delayed-type hypersensitivity (DTH) response toward leishmania antigen in a progressive infection. BALB/b (H-2b) mice showed progressive enhancement in the DTH response, whereas BALB/c (H-2d) mice showed strong DTH at the onset which gradually disappeared (defined as DTH-negative phase) and reappeared again at the later stage of infection. Adoptive transfer of enriched populations of splenic T cells from infected BALB/c mice together with parasite antigen into the footpad of syngenic normal recipients produced a dramatic enhancement in the DTH response, except at the onset of the DTH-negative phase. These observations indicate that adherent cells have a role in suppression of the cell-mediated immune response and also that another mechanism operates at the onset of the DTH-negative phase. This DTH-negative phase was not caused by depletion of DTH-mediating cells from the repertoire, but rather by suppression mediated by a subset of T cell evolved in the course of infection. Characterization on the basis of lymphokine production of the T cells mediating the DTH response and of T cells mediating suppression of the DTH response showed them to be of Th1 and Th2 type, respectively. Studies also indicated that at the onset and the later stages of infection suppression was mediated by adherent cells, but at the onset of DTH-negative phase, in particular, suppression was mediated by Th2 cells. Furthermore, experiments also showed that adherent cells from infected mice gained another property, that of driving B cells, in a T cell-dependent manner.     

Genetic Susceptibility to the Induction of Murine Experimental Autoimmune Orchitis (EAO) without Adjuvant. II. Analysis on Susceptibility to EAO Induction Using F1 Hybrid Mice and Adoptive Transfer System

In our novel murine model of experimental autoimmune orchitis (EAO) induced by two or three injections of viable syngeneic testicular germ cells (TC) alone, significant differences in susceptibility to the induction of EAO were found, and the disease susceptibility did not seem to be associated with a particular H-2 haplotype. The H-2 identical background disparate (highly susceptible × low susceptible)F1 hybrids, (C3H/He × C3H/HeJ)F1 and (C3H/HeJ × C3H/He)F1 mice, were highly susceptible to the induction of EAO. The H-2 identical background disparate (highly susceptible × resistant)F1 hybrids, (C3H/He × C3H/BiKi)F1 and (C3H/BiKi × C3H/He)F1 mice, were low susceptible to the induction of EAO. Both the H-2 and background disparate (highly susceptible × resistant)F1 hybrids, (C3H/He × DBA/2N)F1 and (DBA/2N × C3H/He)F1 mice, were equally resistant to EAO induction. In the susceptible hybrids, both delayed footpad reaction (DFR) and antibody responses to TC increased. On the other hand, in the resistant hybrids, the levels of anti-TC antibodies were elevated but the DFR to TC remains depressed. This suggests that the antibody production and induction of DFR may be under different genetic controls and that cellular immunity plays an important role in this EAO induction.In order to search for the mechanistic basis for low susceptible C3H/HeJ and resistant C3H/BiKi mice, these mice received orchitis-inducible spleen cells (SPCs) from C3H/He mice. C3H/HeJ mice were highly susceptible to passive EAO. In contrast, disease-resistant C3H/BiKi mice failed to develop passive EAO. In addition, we examined whether or not regulatory cells capable of preventing the disease induction were generated in low susceptible C3H/HeJ and resistant C3H/BiKi mice immunized with TC. Transfer of SPCs from TC-immunized C3H/HeJ and C3H/BiKi mice into C3H/He mice before the EAO challenge had no suppressive effect on subsequent disease induction.     

Cutaneous Delayed Type Hypersensitivity in SCID Mice Adoptively Transferred with Lymphocytes is B Cell Independent and H-2 Restricted

Severe combined immunodeficiency (SCID) mice are largely devoid of functional T and B lymphocytes but have normal antigen presenting cell (APC) capacity. The authors have examined the requirements for cutaneous delayed type hypersensitivity (DTH) in SCID recipients by i.p. transfer of freshly isolated naive mature T cells or non-fractionated spleen cells of H-2 compatible or incompatible origin. Recipient SCID mice were epicutaneously sensitized with oxazolone (OXA) within 24 h after cell transfer. SCID mice injected with as few as 10⁵ H-2 compatible BALB/c (H-2^d) spleen cells were able to mount DTH ear swelling reaction upon sensitization and challenge with OXA. Non-fractionated spleen cells from H-2 incompatible B6 or NZW mice were also able to restore DTH capacity in SCID recipients. However, when thymocytes were transferred, only donor mice expressing H-2^d, but not H-2^b and H-2^z, haplotype restored DTH reactivity. Serum levels of IgM and IgM anti-OXA antibodies in SCID mice 27 days post-transfer with 10⁷ BALB/c spleen cells were similar to that of intact donor mice. In contrast, specific antibodies of IgG isotype were approximately only one-tenth of that found in BALB/c controls. The results of this study show that for the development of cutaneous DTH, in SCID mice transferred with T cells, H-2 restricted APC-T cell interaction is required, whereas B cells are not mandatory. Also, SCID mice transferred with splenocytes show signs of defect immunoglobulin switch function. The authors believe that this model of DTH will be useful in delineating the effects of immunomodulatory substances in vivo on distinct host versus donor cell populations.     

Hapten-specific responses to the phenyltrimethyl-amino hapten. I. Evidence for idiotype-anti-idiotype interactions in delayed-type hypersensitivity in mice

The delayed-type hypersensitivity (DTH) reaction to the phenyltrimethylamino (TMA) hapten in mice has been investigated. TMA-derivatized syngeneic spleen cells (TMA-SC) administered s.c. in several strains of mice consistently evoked DTH reactivity, as measured by footpad swelling after challenge with the diazonium salt of TMA. DTH could be induced by low levels of anti-idiotypic antisera (anti-Id) in lieu of antigen. The DTH reaction induced by either mode was hapten-specific, could be transferred into naive recipients by viable lymph node cells but not with serum from immune mice and was not influenced by cyclophoshamide pretreatment. Unlike TMA-SC which induced DTH in all of the strains of the mice tested, anti-Id induced DTH only in strains of the Igh-1e allotype. Positive DTH reactions were induced by anti-Id in the C57.Ige strain (H-2b, Igh-1e) but not in its allotype-congenic partner C57BL/6J (H-2b, Igh-1b). Interestingly this reaction could be suppressed if relatively high amounts of anti-Id were inoculated i.v. just prior to antigen challenge. In addition, the administration of anti-Id 1 h prior to antigen challenge in TMA-SC-sensitized mice significantly blocked the DTH reaction only in the Igh-1e strains. These results demonstrate that the induction and abrogation of TMA-specific DTH by anti-Id is linked to the IgCh locus.     

Anterior chamber inoculation of splenocytes without Fas/Fas-ligand interaction primes for a delayed-type hypersensitivity response rather than inducing anterior chamber-associated immune deviation

The inoculation of antigens into the anterior chamber (AC) of the eye induces an antigen-specific immune response that inhibits delayed-type hypersensitivity (DTH). This regulatory response is known as anterior chamber-associated immune deviation (ACAID). The ACAID response appears to be complex, as it can be elicited by a wide variety of soluble and cell-associated antigens, including foreign, self, tumor, and alloantigens. To evaluate the contribution of Fas/Fas ligand (FasL) interaction to the induction of ACAID to alloantigens, gld and lpr mutant mice were used in conjunction with normal C3H, MRL, and BALB/c mice. ACAID was induced by inoculation of non-irradiated splenocytes from donor mice into the AC of various recipients. After 1 week, recipients were primed intradermally with donor splenocytes. One week later DTH was measured by ear swelling. C3Hgld mutants lacking functional FasL did not develop ACAID after the AC inoculation of BALB/c splenocytes. Conversely, the AC inoculation sensitized these mutants. MRLlpr mutants, which lack Fas, developed ACAID following inoculation of BALB/c cells. AC inoculation of lpr splenocytes did not induce ACAID, but sensitized C3H recipients. Treatment of the AC inoculum with an anti-Fas antibody blocked ACAID induction in a transient manner, as the recipients developed ACAID later. These results show that interaction of the Fas and FasL is required to induce ACAID to allogeneic cells. In the absence of Fas expression on donor splenocytes, or FasL expression by the recipient, AC inoculation primes for a DTH response rather than inducing ACAID.     

Dendritic cells and T cells transfer sensitization for delayed-type hypersensitivity after skin painting with contact sensitizer

The cells involved in the development of delayed-type hypersensitivity (DTH) to the contact sensitizer fluorescein isothiocyanate (FITC) were examined. Cells used for transferring sensitization were obtained from donor mice up to 5 days following skin painting with FITC. Recipient mice were sensitized by footpad injections of dendritic cells (DC) obtained from donor lymph nodes up to 3 days following skin painting, when DC expressed high levels of antigen. DTH, assessed by ear swelling 24 hr following ear challenge with FITC, was detected when recipient mice were challenged 5 days after transfer of DC, but not when ear challenged immediately after transfer. Removal of donor DC using the cytotoxic antibody 33D1, plus complement, either from the DC-enriched population or from whole lymph node cells 24 hr after skin painting, abolished the capacity to transfer DTH. Purified T lymphocytes obtained from donor mice between Days 3 and 5 after skin painting, transferred DTH when recipients were ear challenged with FITC 5 days after footpad injections. DTH also occurred in mice ear challenged with antigen immediately after receiving footpad injections of either normal or irradiated T cells obtained from donors 4 days after skin painting. B cells and macrophages did not transfer sensitization for DTH throughout the time-course. Therefore an early stage in the immune response, where antigen-bearing DC initiated DTH, was distinguished from a later stage, where T cells transferred sensitization.     

T-Cell Effector Function and Unresponsiveness in the Murine Lymphocytic Choriomeningitis Virus Infection

An increase in the virus dose from 10(2) LD50 (low dose) to 10(4) LD50 (high dose) of lymphocytic choriomeningitis virus (LCMV) results in markedly delayed virus clearance, in spite of a potent cytotoxic T-cell (TC) response. However, virus-specific delayed-type hypersensitivity (DTH) reactivity is markedly depressed in high-dose mice, suggesting an association between DTH and virus clearance. When virus-primed memory cells are transferred, DTH reactivity as well as virus-clearing capacity is restored in high-dose mice, indicating that the virus is not present in a changed or concealed form. The role of T-cells mediating DTH (TD cells) in virus clearance was also studied by adoptive transfer to naive recipients. Here the high-dose primed cells did mediate virus clearance, although no DTH reaction was detectable 24-72 h after transfer. However, when footpad swelling was measured 96 h or more after transfer a DTH response emerged, indicating that TD priming had taken place in high-dose animals. Pre-irradiation of high-dose primed cells markedly inhibited the antiviral activity as well as DTH, suggesting that upon transfer to naive recipients TD precursors from high-dose mice would proliferate into effector cells capable of mediating both functions. Treatment with anti-Lyt2+C' abrogated the capacity to induce virus-specific DTH, thus confirming that the cells involved are not helper T (TH) cells. We conclude that the DTH unresponsiveness in high-dose mice reflects a defective differentiation of TD precursor into effector cells which is reversible upon transfer to a less antigen loaded environment. Furthermore, it is suggested that TD function is crucial to the process of virus clearance.     

Augmentation of Natural Killer Cell Activity by Anti-Host Delayed-Type Hypersensitivity during the Graft-versus-Host Reaction in Mice

During a graft-versus-host (GVH) reaction in unirradiated F1 hybrid mice there is a generalized activation of natural killer (NK) cells. We have examined whether the enhanced NK activity is due to an F1 resistance mechanism directed at the parental cells used to induce the GVH reaction. Spleen cells of C57BL/10 origin induce much more NK cell activation in B10F1 hybrids than the opposite parental type, despite a similar intensity of systemic GVH reactions. However, this does not correlate with in vivo resistance of mice with GVH reaction to a local challenge dose of B10 cells. NK cell activation in (CBA X BALB/c)F1 mice with GVH reaction involves both host and donor cells and is preceded by an anti-host delayed-type hypersensitivity (DTH) response. B10 cells have a greater ability to induce DTH in B10F1 mice than cells from the opposite parent. We propose that NK cells are one group of non-specific effector cells recruited by DTH in a GVH reaction and may contribute to the tissue pathology.     

Host defenses in experimental rickettsialpox: genetics of natural resistance to infection.

The genetic basis for natural resistance to lethal infection with Rickettsia akari was studied in over 25 inbred strains, inbred hybrids, and outbred stocks of mice. Inbred mice infected intraperitoneally with the Kaplan strain of R. akari demonstrated three levels of response, susceptible (C3H/HeJ), intermediate (A/HeJ, A/J, A/WySn, BALB/cDub, BALB/cJ, and SJL/J), and resistant (AKR/J, AL/N, BALB/cAnN, BALB/cNCr1BR, C3H/HeN, C57BL/6J, C57L/J, CBA/J, DBA/2J, and SWR/J). No correlation was evident between the six H-2 haplo-types tested and susceptibility to Kaplan infection. Four outbred mouse stocks, Dub: (ICR), Wrc:(ICR), Caw:(CF1), and Mai:(S) were all resistant. The F1 inbred hybrids of resistant X resistant (AKD2F1/J), resistant X intermediate (CB6F1/U), intermediate X intermediate (CAF1/J), and resistant X susceptible (C3D2F1/J) parents were all resistant. The F2 and parental backcross generations of C3H/HeJ and DBA/2J hybrids yielded ratios of resistant to susceptible mice that suggested resistance was under multigeneic control. Susceptible mice (C3H/HeJ) were capable of mounting an immune response, since prior infection with the avirulent Hartford strain of R. akari rendered them resistant to subsequent lethal challenge with the Kaplan strains.     

Induction of delayed-type hypersensitivity to Leishmania major and the concomitant acceleration of disease development in progressive murine cutaneous leishmaniasis.

BALB/c mice injected intradermally with 10(5) or higher doses of formaldehyde-fixed promastigotes (FFP) of Leishmania major developed strong delayed-type hypersensitivity (DTH) to leishmanial antigens injected into the hind footpad 3 to 10 days later. The DTH peaked 15 to 18 h after footpad injection and disappeared by 48 h. This specific DTH correlated with the homing of 51Cr-labeled syngeneic bone marrow cells and the infiltration of proliferating cells to the site of antigen administration. Spleen cells from FFP-sensitized mice also gave significant proliferative response to FFP in vitro. The DTH was adoptively transferable by Lyt-1+2-L3T4+ T cells and was H-2 restricted. DTH could be substantially enhanced by pretreatment with cyclophosphamide or pertussigen. Such DTH enhancement was accompanied by concomitant exacerbation of disease progression after L. major infection. Mice injected intravenously with FFP developed substantial immunity to cutaneous leishmaniasis but specifically suppressed DTH reactivity. Treatment of mice with pertussigen before intravenous immunization, however, abolished the protection and reversed the suppression of DTH. These results therefore demonstrate that the early-appearing type of DTH is not involved in host protection but that it actually facilitates disease progression in cutaneous leishmaniasis. Further evidence, which also shows the nonspecific nature of this disease exacerbation, is provided by local cell transfer experiments. Splenic T cells from mice sensitized to keyhole limpet hemocyanin or FFP induced significantly larger lesions compared with normal T cells when they were transferred into the footpad together with specific antigen and L. major promastigotes.     

MURINE CUTANEOUS LEISHMANIASIS: RESISTANCE IN RECONSTITUTED NUDE MICE AND SEVERAL F1 HYBRIDS INFECTED WITH LEISHMANIA TROPICA MAJOR

After cutaneous injection of promastigotes of an isolate of the intramacrophage protozoan parasite, Leishmania tropica major, mouse strains develop chronic cutaneous lesions or show a resolving pattern of disease. On this basis, they can be classified as resistant (e.g. CBA/H and C57BL/6) or susceptible (e.g. BALB/c, BALB/c.H-2^b and BALB/c.H-2^k). Hypothymic nude (nu/nu) mice of either BALB/c, CBA/H or C57BL/6 genotype are susceptible to chronic disease. However, nude mice of these genotypes, including BALB/c, are resistant to chronic cutaneous leishmaniasis when injected at the time of parasite challenge with small numbers of H-2 compatible lymphoid cells from normal mice. Nude mice remain susceptible when injected with fully H-2 incompatible cells. Using cells from H-2 mutant mice for reconstitution of resistance in C57BL/6.nu/nu mice, evidence was obtained that I region compatibility is necessary for cells to mediate host-protective effects. Cells from chronically-diseased BALB/c mice do not have protective effects in BALB/c.nu/nu mice at any cell dose and will abrogate the resistance-promoting effect of lymphoid cell populations from chronically-diseased BALB/c.H-2^k and BALB/c.H-2^b mice can be demonstrated when assayed at certain cell doses in H-2 compatible CBA/H.nu/nu and C57BL/6.nu/nu mice, respectively. The data suggest that chronically-diseased (genetically-susceptible) mice contain a mixture of resistance-promoting and disease-promoting T cells in their peripheral lymphoid organs and that expression of the resistance-promoting subset can occur in nude mice of resistant genotype. Previous data have indicated that Lyl⁺2^? T cells are efficient mediators of both T cell-dependent activities. No evidence for the operation of disease-promoting or resistance-promoting antibodies in perpetuation or resolution of disease has been obtained in extensive serum transfer experiments. Some discrepancies exist in the literature on the question of the dominance of susceptibility or resistance in F₁ hybrid mice. A re-examination of susceptibility/resistance in F₁ hybrids between BALB/c and several other parental strains was undertaken using cloned pathogenic promastigotes derived from a heterogeneous L. t. major isolate in order to reduce effects of parasite heterogeneity in the analysis. Resistance was dominant in some but not all F₁ hybrids, with most showing a delayed healing pattern of disease relative to the resistant parental strain. Despite the use of genetically-homogeneous parasites, the analysis was complicated by variability within groups of F₁ hybrid mice as well as between males and females and between F₁ hybrids of reciprocal crosses. A hypothesis based on antigen and H-2 expression on infected macrophages is advanced to account for the balance between the effects of resistance-promoting and disease-promoting Lyl⁺2^? T cells in mice of various genotypes.     

Immune response against the T-independent antigen alpha (1----3) dextran. II. Occurrence of B gamma memory cells in the course of immunization with the native polysaccharide is T cell dependent

The immune response of BALB/c mice against the bacterial antigen dextran B1355S [alpha (1----3)dextran] (Dex), a class 2 T cell-independent antigen, is largely restricted to IgM class antibody production. However, despite the fact that Dex fails to give rise to enhanced IgG responses upon repeated immunization, the development of Dex-specific B gamma memory cells, i.e. resting B cells committed to the production of specific IgG antibodies, is observed upon immunization with Dex. These B gamma memory cells, albeit not activated to IgG production in situ, can be activated upon adoptive transfer into irradiated congenic BALB Ighb mice. This mouse strain is a nonresponder strain to Dex. The expression of adoptively transferred Dex-specific B gamma memory cells is T cell-independent as T cell depletion of spleen prior to cell transfer into BALB.Ighb recipients does not abolish the IgG response. Dex-primed athymic BALB/c nude mice, in contrast to euthymic mice, give a pronounced primary IgG response but do not develop Dex-specific B gamma memory cells. Yet, they do so when reconstituted with syngenic T cells prior to immunization. This indicates that the formation of Dex-specific B gamma memory cells requires the presence of functional T cells. The pronounced primary IgG anti-Dex response of nude mice is greatly impaired by T cell reconstitution. Thus, with regard to T cell dependence, there is an inverse relationship between the formation of B gamma memory cells and the capacity to produce IgG anti-Dex. Dex-specific B gamma memory cells from BALB/c mice are expressed in congenic BALB.Ighb recipients (nonresponder to Dex) but not when transferred into identically treated syngenic hosts. This also applies to memory cells from Dex-primed female (CBA/N X BALB/c)F1 [NBF1] or from (BALB/c X CBA/N)F1 hybrids. Dex-specific B gamma memory cells from these donors are demonstrable upon adoptive transfer into BALB.Ighb mice, but they are not expressed when transferred into syngenic recipients, including male NBF1 hybrids. NBF1 males, albeit possessing the VH-Dex+ allele, do not mount humoral responses to Dex, a deficiency which is ascribed to an X chromosome-linked B cell defect. The apparent absence of Dex-specific antibodies in NBF1 males provides an opportunity to examine whether B gamma memory cell expression is inhibited in syngenic recipients by anti-Dex or autologous anti-idiotype antibodies.(ABSTRACT TRUNCATED AT 400 WORDS)     

Genetic studies of the murine corneal response to Pseudomonas aeruginosa.

The murine genetic control of resistance to Pseudomonas aeruginosa eye infection previously has been demonstrated to be regulated by two complementing dominant genes, PsCR1 and PsCR2. The PsCR1 locus apparently is not associated with the H-2 complex, whereas the PsCR2 locus could not definitively be associated with H-2. In this study we attempted to demonstrate a possible H-2 linkage of the PsCR2 locus. A panel of inbred congenic strains varying with either the H-2 haplotype or genetic background from inbred partners of C57BL/10, C3H, A, and BALB/c strains were characterized for their P. aeruginosa infectivity phenotypes. These studies indicated that the PsCR2 locus is not associated with the H-2 locus. Furthermore, variations of the H-2 haplotype did not change the resistance patterns observed in these strains. However, BALB.B and BALB.K congenic lines were resistant to P. aeruginosa eye infection, whereas BALB/cJ mice were susceptible. Examination of hybrids (BALB.K X BALB/cJ)F1 and (BALB.B X BALB/cJ)F1 demonstrated that an autosomal dominant gene(s), PsCR, confers resistance. Segregation analysis for the H-2 haplotype and the PsCR gene in offspring of backcross matings with the BALB/cJ parental strain suggested that this PsCR gene is not linked to the H-2 complex and has an inheritance pattern of a single locus or several tightly linked loci.     

Secondary delayed-type hypersensitivity to sheep red blood cells and minor histocompatibility antigens in mice: transfer of memory by recirculating thoracic duct lymphocytes

Secondary delayed-type hypersensitivity (DTH) in mice to sheep red blood cells (SRBC) and minor histocompatibility (H) antigens is dependent on long-lived memory T cells. In this paper we investigated whether these memory T cells recirculate. It was shown that late phase "immune' thoracic duct lymphocytes (TDL) from mice which were immunized with SRBC or non-H-2-incompatible spleen cells several weeks previously could adoptively transfer secondary DTH to these antigens. Passing the immune TDL through intermediate recipients demonstrated that these SRBC- or minor H-antigen-reactive memory T cells recirculate from blood to lymph. In contrast to mice immunized with minor H antigens, no secondary type DTH reactivity could be demonstrated in mice immunized with H-2-incompatible spleen cells. Also, after adoptive transfer of TDL from mice immunized with H-2-alloantigens, it was impossible to demonstrate an accelerated DTH reactivity.     

Dominant suppressive effect of the silent Eb alpha allele on an in vivo T helper cell response under Ed beta Ed alpha region-linked immune response gene control

Previous adoptive spleen cell transfer experiments have demonstrated that an immune response (Ir) gene linked to the Ed beta Ed alpha region allows BALB/c T helper lymphocytes (Th) to respond to an idiotope on the V lambda 2(315) fragment of isologous myeloma protein M315. BALB.K (H-2k) and BALB.B (H-2b) do not respond to V lambda 2(315). While (H-2d X H-2k)F1 hybrids have been shown to be responders, it is now demonstrated that (H-2d X H-2b)F1 hybrids are low responders. By crossing BALB/c with various H-2 recombinants on B10 background and probing Th responsiveness to V lambda 2(315) in these F1 hybrids, the dominant suppressive gene of the H-2b haplotype is mapped to Eb alpha Sb. It is argued that the suppressive gene is Eb alpha, which is a silent allele. A likely explanation for the suppressive effect of the Eb alpha allele is that reduced amounts of Ed beta: Ed alpha restriction elements are present on antigen-presenting cells of (H-2d X H-2b)F1 hybrids because only one E alpha gene is functional in such mice. The present report extends previous in vitro findings from other laboratories to the in vivo situation and suggests that silent alleles for class II molecule chains may profoundly affect certain immune responses of individuals heterozygous for the silent allele.     

In vitro induction of suppressor T-cells in delayed-type hypersensitivity to BCG and an essential role of I-J positive accessory cells

Antigen-specific suppressor T-cells in delayed-type hypersensitivity (DTH) to BCG were induced in vitro. Normal spleen cells of C3H/He mice were incubated with 50 micrograms of PPD per ml for 4 days at 37 degrees C, and the non-adherent cells in the culture were transferred intravenously into cyclophosphamide (CY)-treated syngeneic recipients. The recipients were immunized to BCG immediately after the cell transfer, and DTH was measured by the footpad reaction to PPD two weeks later. Footpad reaction to PPD was positive in CY-treated C3H/He mice immunized to BCG, while it was suppressed by the transfer of the in vitro induced suppressor cells. When the suppressor cells were treated with anti-thy-1.2 antiserum and complement before transfer, the suppression was abrogated. Next, the spleen cells were separated into plastic adherent and non-adherent fractions. After treatment with anti-thy-1.2 and complement, the adherent cells were treated with either anti-I-Jk or anti-I-Ak antiserum and complement. Then, they were reconstituted with the non-adherent cells and cultured with PPD. Treatment of the adherent cells with anti-I-Jk antiserum and complement abrogated the suppressor cell induction, while the treatment with anti-I-Ak had no effect. These facts indicate that I-J positive non-T-adherent cells play an essential role in the induction of suppressor cells in DTH.     

The production of contact sensitivity by the injection into the footpads of recipients of the lymph node cells from mice 1 day after painting the skin with contact sensitizing agent: requirement for matching at the major histocompatibility complex between donor and recipient mice.

Donor mice were painted on the skin of the abdomen with the contact sensitizing agent, oxazolone. One day later 2-5 x 10(6) cells from the regional lymph nodes were injected into the footpads of recipient mice. Contact sensitivity was detected 6 days later by challenging the ears of the recipients and measuring the increase of thickness at 24 h. Good contact sensitivity was obtained when CBA cells were injected into CBA mice and BALB/c cells injected into BALB/c mice; the injection of BALB/c (H-2d) cells into CBA (H-2k) mice and vice versa failed to give rise to contact sensitivity. Hybrid F1 cells gave intermediate responses. The contact sensitivity caused by the injection of small numbers of lymph node cells into the footpad is interpreted as a mode of active immunization and the present results show that this only occurs when there is genetic matching at the major histocompatibility complex between the donor and the recipient mouse.