Tail Skin Grafts Research Articles

Acceptance of class II major histocompatibility complex disparate skin grafts associated with suppressor cells and elevated langerhans cell numbers

Class II major histocompatibility complex (MHC) molecules are only present on Langerhans cells (LC) in normal murine epidermis. Depletion of this antigen with the chemical carcinogen dimethylbenzanthracene (DMBA) causes I-E disparate B10.A(2R) congenic tail skin to be accepted permanently when grafted onto B10.A(4R) recipients. Adoptive transfer of spleen cells from these recipients into naive syngeneic hosts inhibited the ability of the host mice to reject untreated B10.A(2R) tail skin grafts. Hence DMBA-treated LC depleted I-E disparate skin grafts activate suppressor cells which did not inhibit BALB/c mice from rejecting a B10.A(2R) tail skin graft. In contrast, the tobacco derived carcinogen benzo(a)pyrene (BP) increased the number of epidermal LC but had no effect on either class I or class II MHC disparate skin graft survival time. This confirms that the number of class II MHC-positive LC is critical for the initiation of skin graft rejection; when the threshold level is attained graft rejection proceeds at a maximal rate that cannot be enhanced by raising the number of LC. The tolerant skin grafts had increased numbers of LC; this was not observed in syngeneic grafts and therefore may be related to the active suppression of immunity.

Cellular immune response of a marsupial, Monodelphis domestica

Marsupials are interesting subjects for studies of comparative and developmental immunology because they separated from eutherian mammals over 100 million years ago and because the newborns are still in a fetal state. We studied cellular immunity in a fully pedigreed colony of the marsupial, M. domestica (commonly called the gray short-tailed opossum). Peripheral blood lymphocytes were separated on nylon wool columns into adherent cells bearing surface immunoglobulin (B cells) and nonadherent cells (T cells) recovered in the ratio of 1:3. Peripheral blood lymphocytes responded by proliferation to Con A and other mitogens. Nonadherent cells were responsive to Con A, but adherent cells were not. Peripheral blood lymphocytes were stimulated weakly or not at all by allogeneic or xenogeneic (mouse) cells in mixed lymphocyte culture. Despite the weak MLC response, which was not due to genetic homogeneity, allogeneic and xenogeneic tail skin grafts were rejected promptly. These data suggest that the cellular immune response of M. domestica is similar to that of eutherian mammals with the notable exception of weak MLC responses

Biochemical analysis of related, independently arising histocompatibility mutants: bm17 and KB-98 enlarge the “bg series” of H-2Kb mutants

The "bg" series of MHC mutations is the most prevalent type of mutations of Kb in C57BL/6 mice screened by reciprocal tail skin grafting. The basis for identification of this series of mutations is the incompatibility of grafts between the parental B6 and the mutant. This series takes the longest to reciprocally reject the skin grafts. The series can be subdivided into "bg 1" and "bg 2" groups based on Kb-restricted recognition of virus-infected mutant target cells. The biochemical basis for these mutations are amino acid substitutions at residues 116 and 121 of the Kb transplantation antigen. These substitutions do not alter monoclonal antibody binding sites. The structural basis of MAb binding and the genetic basis of the mutation are discussed.

In vivo induction of antigen-specific transplantation tolerance to Qa1a by exposure to alloantigen in the absence of T-cell help.

A goal of transplantation immunology is to be able to induce antigen-specific tolerance in transplant recipients. In the present study we describe an in vivo model of antigen-specific transplantation tolerance to skin allografts using mice congenic at Qa1, a ubiquitously expressed class I-like molecule encoded to the right of H-2D. B6 mice are deficient in Qa1a-specific T-helper cells and only reject Qa1a disparate tail skin grafts when a second graft expressing additional helper determinants is also present. We report that animals initially engrafted with Qa1a disparate skin, in the absence of any source of additional help, are rendered tolerant to Qa1a disparate skin allografts despite the subsequent presence of inducer skin grafts expressing additional helper allodeterminants. The nonresponsive state is Qa1a-specific, because HY-bearing inducer grafts are rejected normally. In vitro, Qa1a-tolerant animals are specifically unable to generate anti-Qa1a T-killer cells, which provides the cellular basis for their failure in vivo to reject Qa1a skin allografts. Thus, initial exposure to Qa1a allodeterminants, in the absence of T-cell help, leads to a state of Qa1a-specific transplantation tolerance. This study suggests that antigen-specific transplantation tolerance may be induced by exposing naive T-killer cells to tissue alloantigens under conditions in which T-cell help is not generated.

IMMUNOLOGICAL UNRESPONSIVENESS INDUCED BY ULTRAVIOLET-B-IRRADIATED AND NONIRRADIATED SKIN GRAFTS

We previously reported that ultraviolet-B-irradiated B10.AQR tail skin grafts were permanently accepted by B10.T6R recipients in about half the cases. Such a beneficial effect on graft survival could only be demonstrated in this particular combination. We have now investigated whether these animals had become tolerant to the donor strain antigens. Nonirradiated B10.AQR tail skin grafted 50 days after acceptance of a UVB-irradiated B10.AQR graft was accepted in 9/9 cases, indicating that these animals had become tolerant to the B10.AQR alloantigens. However, secondary grafts on animals that had rejected the first graft also showed a prolonged or definite survival. This tolerance was specific; B10.T6R mice tolerant to B10.AQR grafts rejected B10 skin grafts, while F1(B10A X B10.AQR) and F1(B10 X B10A) grafts, sharing class II antigens with B10.AQR, had a slightly prolonged graft survival. Cells of tolerant animals showed normal proliferative responses against B10.AQR antigens. However, when autologous serum was added, proliferation was specifically suppressed. Likewise, this specific tolerance could be transferred with serum or serum and cells but not with cells only. Analysis of the sera of these animals showed long-lasting and donor-specific high-titered cytotoxic antibody titers, which are likely to play a pivotal role in the observed suppression.

Induction of allograft tolerance to the H-Y antigen in adult C57BL/6 mice: Differential effects on delayed-type hypersensitivity and cytolytic T-lymphocyte activity

This study describes the induction of allograft tolerance to the “male-specific,” minor histocompatibility antigen, H-Y, in adult C57BL/6 female mice, and the effects of this tolerance induction on two immune parameters associated with graft rejection: delayed-type hypersensitivity (DTH) and cytolytic T-lymphocytes (CTL). B6 females tolerized to H-Y, by a single iv injection of C57BL/6 male lymphocytes, exhibited prolonged or permanent survival of B6 male tail skin grafts. Graft-induced DTH against H-Y antigen was reduced or abrogated in tolerized females. Delayed onset of graft rejection in partially tolerant females correlated with delayed onset of DTH, and eventual rejection of grafts was accompanied by an increase in H-Y-specific DTH. In contrast, H-Y-specific CTL activity was not consistent with graft status. These data demonstrate a correlation between H-Y-specific DTH and rejection of male skin grafts by B6 female mice and are most consistent with a major effector role for DTH in chronic graft rejection.

In vivo and in vitro characterization of specific hyporeactivity to skin xenografts in mixed xenogeneically reconstituted mice (B10 + F344 rat----B10).

Mixed xenogeneically reconstituted mice (F344 rat + C57BL/10Sn----C57BL/10Sn), which specifically retain F344 tail skin xenografts, were studied for the specificity of such hyporeactivity and for in vitro reactivity and immunocompetence. Survival of mixed reconstituted animals was excellent, without evidence for graft vs. host disease. Donor-type tail skin grafts were specifically prolonged (mean survival time = 80 d) in comparison with normal controls and syngeneically reconstituted animals. In vitro, such animals manifested specific hyporeactivity by mixed lymphocyte reaction and cell-mediated lympholysis to F344 rat and B10 cells, with normal response to third-party rat (Wistar-Furth) and mouse (B10.BR). Examination of lymphoid tissues with a fluorescence-activated cell sorter revealed low levels, if any, of donor-type cells detectable. This system offers a model for investigation of xenogeneic transplantation tolerance.

Tailskin grafts do not show accelerated rejection on splenectomized hosts

Tailskin grafts do not show accelerated rejection on splenectomized hosts

Genetic mapping and immunogenetic characterization of theH- 2fb mutation in the mouse

Rejection of tailskin grafts exchanged between two male hybrids of the cross B10.M × B10.RIII(71NS) revealed a mutation in theH-2 ( f ) haplotype from the B10.M congenic line. Complementation studies with skin grafting and cell-mediated lympholysis showed the mutant, namedH-2 ( fb ), to be different from anotherH-2 ( f ) mutant,H-2 ( fa ), and further, that the HH-2 ( fb ) mutation occurred in theD end of theH-2 complex. Reciprocal skin grafts exchanged between mutant and normal mice were rejected. Hemagglutinating antibody reactive with B10.M cells was raised in the mutant mice. Mutant spleen cells responded weakly, but significantly, to wild-type cells in a mixed lymphocyte culture and in a graftversus-host assay, but no response was seen in the opposite direction. However, cytotoxic effector cells were generated against target cells in both directions in a cell-mediated lympholysis assay.

Apparent Lack of H-2 Restriction of Allograft Rejection

BALB/c ByJ (H-2d) mice immunized with tail skin grafts of either B10.D2 (H-2d) or B10 (H-2b) demonstrated similar second set rejection of B10.D2 tail skin. This apparent lack of H-2 restriction was not due to the induction of a new population of cytotoxic T lymphocytes (Tc) since 450R given 24 hr before the challenge graft did not abrogate the second set reactivity. Host macrophage processing or anti-Qa-2 reactivity was also not the explanation for the lack of H-2 restriction since immunization of BALB/c Li mice with either B10.D2 or B10 tail skin grafts resulted in second set rejection of B10 tail skin. Shared public H-2 specificities of H-2d and H-2b may result in cross-reactive Tc, thus causing the apparent lack of H-2 restriction. However, no H-2 restriction of allograft rejection is observed when only one public H-2 specificity is shared between the recipient and the allogeneic challenge graft (H-2f and H-2k combination). These results suggest that H-2 restriction of T cell cytotoxicity has little relevance in allograft rejection because 1) one public H-2 specificity is sufficient to cause cross-reactivity or 2) Tc are not the major effectors of allograft rejection.

Marrow allograft success inW/W v anemic mice: Relation to skin graft survival times

Genetically anemicW/W v mice were cured by marrow allografts from donors of 13 out of 18 tested strains that differed at non-H-2 histocompatibility alleles defined by skin or tumor grafting. They were also cured by donors from all four tested congenic lines whose antigenic differences had been defined by induction of serum antibodies. They were not cured acrossH-2 differences. Tail skin graft survival times on uncuredW/W v recipients were determined for all congenic lines used as marrow donors. The longest and shortest skin graft survival times predicted correctly marrow graft success or failure. NoW/W v mice were cured by marrow grafts from donors of the three congenic lines whose skin grafts were rejected in fewer than three weeks. Almost everyW/W v mouse grafted was cured by marrow grafts from donors of the 13 congenic lines whose skin grafts survived longest, from 11 to more than 25 weeks. Intermediate skin graft survival times failed to predict whether marrow grafts would succeed.W/W v mice were cured by marrow from four congenic lines with mean skin graft survival times of 4.2, 4.4, 8, and 9 weeks, while marrow grafts failed from other congenic lines with mean skin graft survival times of 3.3, 3.4, 4.8, and 8.7 weeks. The simplest explanation for these results is that the antigens specified by theH-2, H-3, H-4, H-25, andH-28 loci are strongly immunogenic on both marrow precursor cells and skin,H-17 andH-24 are strongly immunogenic on skin but not on marrow, andH-12 is strongly immunogenic on marrow precursor cells but less strongly on skin.

Genetics of histocompatibility in mice

Twenty-five new congenic lines with distinctive BALB/cBy-strain histocompatibility alleles introduced onto the C57BL/6By-strain background by a regimen of backcrossing and tailskin grafting have been established. Twenty-one of the histocompatibility loci represented by these lines are new, while four duplicate theH-1, H-2, H-7, andH-8 loci identified by Snell.

The induction of long-term immunological tolerance

CBA mice were rendered tolerant by a prolonged course of antilymphocyte serum and the subsequent injection of allogeneic or xenogeneic lymphoid cells. Control mice received no other therapy. Experimental mice were grafted with allogeneic or xenogeneic lymph nodes prior to the administration of the cellular inocula. The survival pattern of tail-skin grafts was assessed in all mice. Those animals that were grafted with lymph node transplants failed to reject the skin grafts with the same rapidity or pattern as mice that were treated with ALS and cells alone. Furthermore, graft-v.-host disease was not noted in either the allogeneic or xenogeneic chimeras. The possible mechanisms of this durable chimerism are discussed. The experiments are still preliminary and others are in progress to evaluate the survival patterns of lymphoid cells in their heterotopic but syngeneic environment.

Implantation, transplantation, and epithelial-mesenchymal relationships in the rat uterus.

Free tail skin grafts or suspensions of viable epidermal cells have been placed in the atraumatized uterus of isologous rat hosts and allowed to "implant" of their own accord to study the possible uniqueness of this site for other than nature's transplants, i.e. conceptuses, and its response to unnatural grafts. Despite the presence of an intact endometrial epithelium, free skin grafts heal-in rapidly, provided that a state of estrogen excess is established at the time of transplantation. In the absence of estrogen most of the grafts failed to implant. Once established, the grafts survive indefinitely without further estrogen. However, if at any stage a state of continual estrus is established, skin epidermis migrates centrifugally from the graft perimeter invasively replacing the native uterine epithelium. The results of an analysis of the modus operandi of this estrogen-facilitated epidermal migration in utero sustain the view that the hormone acts upon the uterine stroma rather than upon the epidermal cells. When grafts of lingual mucosa or vaginal "skin" were placed in the uteri of rats maintained in chronic estrus, migration of epidermis took place even more vigorously than from tail skin. These epithelia conserved their distinctive histologic specificities indefinitely when growing as heterotypic recombinants on the alien mesenchymal stroma of the uterus. Monodisperse suspensions of epidermal cells appear to "implant" and establish small epidermal plaques in the uterus only at sites predestined to accept conceptuses. That the endocrinologic parameters for the establishment of skin grafts in the uterus are similar to those for blastocysts is suggested by the finding that both kinds of graft can become established in the same uterine horn in the absence of exogenous hormones.

Tolerance of rat skin grafts in adult mice

Abstract Adult CBA mice may be made tolerant of August rat tail skin grafts by short intensive treatments with antilymphocyte serum followed by massive (∼100 × 106 cells) injections of August rat lymphoid cells. Prolonged tolerance can be achieved only in thymectomized mice, and is accompanied by lymphoid cell chimerism and the manufacture of rat protein. The rat lymphoid cell donors must be treated beforehand with antirat ALS to avoid complications associated with graft versus host reactions. Rejection of rat skin grafts is accompanied by the formation of high titres of antirat hemolysins and lymphocytotoxins, but in “tolerant” mice the rat skin survives in the continual presence of antirat antibodies. Under certain circumstances, however, passive transfusion of high titre mouse antirat serum can precipitate the breakdown of rat skin grafts. It is doubtful if this represents the primary mechanism of rejection. The reaction of normal mice against rat skin is essentially an intensified allograft reaction; it is at all events wholly immunological in character.

Tolerance of rat skin grafts in adult mice.

Adult CBA mice may be made tolerant of August rat tail skin grafts by short intensive treatments with antilymphocyte serum followed by massive ( approximately 100 x 10(6) cells) injections of August rat lymphoid cells. Prolonged tolerance can be achieved only in thymectomized mice, and is accompanied by lymphoid cell chimerism and the manufacture of rat protein. The rat lymphoid cell donors must be treated beforehand with antirat ALS to avoid complications associated with graft versus host reactions. Rejection of rat skin grafts is accompanied by the formation of high titres of antirat hemolysins and lymphocytotoxins, but in "tolerant" mice the rat skin survives in the continual presence of antirat antibodies. Under certain circumstances, however, passive transfusion of high titre mouse antirat serum can precipitate the breakdown of rat skin grafts. It is doubtful if this represents the primary mechanism of rejection. The reaction of normal mice against rat skin is essentially an intensified allograft reaction; it is at all events wholly immunological in character.

Histocompatibility of skin grafts from mice of F1, F2, and F3 generations on F1 generation hosts.

Hybrid mice of the F1, F2, and F3 generations were produced from an initial cross of inbred strains C57BL/6 and BALB/c. Orthotopic female tailskin grafts of one F1 donor of 167, two F2 donors of 218, and no F3 donors of 218 were found to be rejected by F1 female hosts. The rejections could be attributed to mutation; otherwise, the results upheld the genetic laws of transplantation.

The effect of graft thickness, donor site and graft bed on graft shrinkage in the hooded rat

Summary 1.Full thickness homografts of body, tail and ear skin, and measured split thickness grafts of body and tail skin, were applied to prepared sites on the chests of homozygous rats. 2.Grafts applied to the deep fascia (as tested with tail skin grafts), shrank slightly less than grafts applied to the panniculus carnosus. 3.The orientation of the graft (as tested with body skin grafts), did not affect the amount of shrinkage; but shrinkage was consistently greater in the plane of the dressing tension. 4.Of the full thickness grafts, these from the body skin shrank most initially and then enlarged to a final 65 per cent. of their original area. Tail skin shrank to about 65 per cent. and remained constant. Ear skin after a variable amount of shrinkage, enlarged to its original size. 5.With body skin, the split thickness grafts shrank more than full thickness, the thinner the graft the more the initial contraction and the less the subsequent enlargement, but equal thickness split skin grafts from the dorsum shrank more than those from the ventrum. 6.With tail skin the split thickness grafts shrank less than full thickness skin. 7.It is concluded that the nature of the donor site as well as the thickness of the graft affect the amount of shrinkage, and that the orientation, and the graft bed (if mobile) have little or no effect.

Inherited histocompatibility changes in progeny of irradiated and unirradiated inbred mice

(1) F1-hybrid mice derived from a cross of the highly inbred strains: C57BL/6 and BALB/c, were tested for inherited changes of histocompatibility by an orthotopic inter-exchange of tail-skin grafts. The fathers of tested mice received either 522 rads of gonadal X-irradiation, or received no irradiation 2 months prior to mating.(2) Thirty-two mice with altered histocompatibilities were found in a total of 2572 complete tests. All of those mutant mice (twenty-one) that produced an adequate number of offspring were shown to pass the incompatibility on to their progeny.(3) Mutants were classified as to whether they effected a gain, a loss or both a gain and a loss in antigen specificity as determined by whether they rejected skin of donor mice or their skin was rejected by host mice. Twenty-six were clearly of the gain type, five were most likely gain type and only one showed both a loss and a gain effect. There was no clearcut evidence that loss types had occurred. The preponderance of gain types was tentatively explained as an artifact of the system used for the assay.(4) Several of the detected mutants were probably from parents carrying mutations that originated in past generations, for some mutant mice occurred in clusters.(5) There was no apparent effect of paternal irradiation (522 rads) on mutation frequency. The induced mutation rate was estimated to be less than 2·6 × 10−5/ gamete/rad.(6) Independent data on isografts from F1hybrids of proven non-carrier pedigreed parents provided an estimate of spontaneous mutation rate of 6·75 × 10−3/ gamete.(7) The estimate of doubling dose (greater than 260 rads) was consistent with the estimates for recessive lethals and visibles in mice.

A search for histocompatibility differences between irradiated sublines of inbred mice

(1) A tail-skin grafting method was used to test for histocompatibility differences between members of thirteen sublines of C3H inbred mice, kept in a 1 r./night gamma radiation field for twelve generations, on the average, and separated from each other by about thirty-four generations.(2) No homograft rejections occurred, so there was no evidence to suggest that mutations at histocompatibility loci had taken place in any of these sublines. Calculations show that this finding does not conflict with the idea that a fairly large number of loci are involved, having mutation rates similar to those already known in the mouse.