Positive H-Y antigen testing in a case of XY gonadal absence syndrome

H-Y antigen testing was positive in a case of pseudohermaphroditism due to XY gonadal absence syyndrome. In conclusion, H-Y antigen may be present even in the absence of testes. Also, the syndrome could not have originated from a defect in the H-Y antigen system.     

45, X Constitution in a H-Y antigen positive boy with partial monosomy 5p

This is the second reported case of a boy with partial monosomy 5p and 45, X constitution. He has the typical clinical features of the Cri-du-chat syndrome and no Turner stigmata. The normal male development of external genitalia in the patient is related to the presence of the H-Y antigen.     

H-Y antigen in 46,XY pure testicular dysgenesis

Clinical, cytogenetic, pathologic and histocompatibility-Y (H-Y) antigen studies were performed on a phenotypic female with primary amenorrhea and streak gonads. Pathological examination of tissues removed at total hysterectomy and bilateral salpingo-gonadectomy showed gonadoblastoma and dysgerminoma of left streak. A single F-body (Y chromosome) was found in buccal smears. Analysis of blood cells and tumor fibroblasts showed a 46, XY chromosome constitution (Q-banding). The data were consistent with a diagnosis of 46, XY pure testicular dysgenesis. Positive results for H-Y antigen were found in this case.     

H-Y antigen generating and receptor systems in abnormal sexual development

This paper reports data from one case of XY gonadal dysgenesis and one case of XY mixed gonadal dysgenesis. Their H-Y antigen expression was determined on peripheral blood leukocytes and on gonad-derived fibroblasts. The former case was found to be H-Y negative while the latter was H-Y positive. The origin of these disorders is discussed in terms of mutations affecting the H-Y antigen generating and receptor systems.     

XY (H-Y+) gonadal dysgenesis

Summary The gonads of 4 phenotypically female individuals with XY chromosomal constitution and signs of virilisation were examined by light microscopy. Electron microscopic examination was also performed in two cases. Serological analysis of H-Y antigen titer yielded positive results. The matrix of the gonads is shown to be ovarian stroma, in which tubular and follicular structures are embedded. The epithelia of the follicles resemble granulosa cells of the ovary, the tubular epithelia are resemble Sertoli cells. Tubules and follicles both show extensive regressive changes. A varying number of Leyding cells/stroma lutein cells were found in each gonad. The different degree of development of testicular and ovarian structures in the dysgenetic gonads might be explained by a defect of the gonadal specific receptor for the H-Y antigen, this defect varying in time of occurrence, duration and severity.     

H-Y typing in the ELISA

In a series of quantitative absorptions, biotin-conjugated monoclonal H-Y antibody was reacted with a plated source of H-Y antigen in the ELISA. H-Y was demonstrated in testis supernatant fluid of the mouse in this system, and in cells from males of the mouse, bovine and human and females of the chicken, thereby confirming its evolutionary persistence, and underscoring its presumptive role in the primary determination of vertebrate sex.     

Presence of H-Y antigen in patients with ullrich-Turner syndrome and X-chromosome rearrangements

Cells from eight of ten patients with gonadal dysgenesis and an isochromosome for the long arm of X, (i(Xq)), have been found to be H-Y antigen-positive, using an assay that employs rat antiserum and Raji cells. In addition, two patients with del(Xq) were also found to be H-Y antigen-positive, whereas four patients in whom only a 45,X line was detected were H-Y antigen-negative. These findings suggest that the X chromosome plays a role in the expression of H-Y antigen in the absence of a Y chromosome. Since our patients with i(Xq) show no evidence of testicular differentiation, it is clear that there is not enough H-Y antigen on these patients' cells to direct the development of a testis. These findings are consistent with the view that the normal functioning of genes on the X and the Y chromosomes is necessary for testicular organogenesis to occur.     

H-Y Antigen Expression Patterns in Human X- and Y-Chromosome-Bearing Spermatozoa

PROBLEM: Restricted expression of H-Y antigen on Y-chromosome-bearing sperm has been reported in some species, although such preferential expression for H-Y antigen in human sperm has yet to be described. In this study, an immunomagnetic approach was used to characterize antigen expression patterns as a function of sex-chromosome content. METHOD OF STUDY: Human sperm was treated with monoclonal immunoglobulin (Ig) M antibodies directed against H-Y antigen. This preparation then was incubated with sheep antimouse IgM antibody affixed to paramagnetic beads, which then were exposed to a magnetic field and sorted. X- and Y-chromosome frequencies in the two subgroups of sperm were assayed by multiprobe fluorescent in situ hybridization (FISH). RESULTS: Sperm were immunomagnetically separated into two populations: a reactive group (presumably, H-Y Ag+); and a nonreactive group (presumably, H-Y Ag-). Triple-color FISH analysis of 1,600 spermatozoa (800 in each group) showed the antigen's expression to be somewhat more prevalent among Y-chromosome-bearing sperm (54.1%), but a large proportion of Y-chromosome-bearing sperm (49.0%) did not express this antigen. The difference was not significant (P = 0.43). CONCLUSIONS: The expression of H-Y antigen has a slightly higher frequency in human sperm containing the Y-chromosome, but its expression among X-chromosome-bearing sperm also is considerable. Current immunologic techniques relying on this antigen are unlikely to effect the sex selection of human sperm.     

Diagnostic applications of H-Y serology: H-Y negative phenotype in cells from 45, X/ 46, XY fetus with testes

Sexual dysmorphism should be considered likely in cases in which H-Y- phenotype and XY complement are found together. In the case described here, a pregnancy was terminated at nineteen weeks of gestation after 45,X and 46,XY cell lines were detected among cultured amniocytes. The fetus was a male with hypospadias and intraabdominal testes containing irregular tubules and hyperplastic interstitium. Cultured skin fibroblasts, containing 45,X and 46,XY lines in ratio of 18:2, were typed H-Y antigen negative. This underscores the danger of predicting gonadal type on the basis of somatic H-Y phenotype.     

Correlation between testicular tissue and H-Y phenotype in intersex patients

H-Y antigen was tested in 13 humans with testicular tissue and disturbed primary and/or secondary sex differentiation. All cases typed H-Y positive. This finding is in agreement with the idea that testicular tissue only differentiates in the presence of H-Y antigen.     

H-Y antigen expression in patients with X-autosomal translocations and gonadal dysgenesis

Cells from three patients with early gonadal failure and a balanced reciprocal translocation involving the long arm of the X chromosome and an autosome were studied. Fibroblasts from a patient with a similar balanced reciprocal translocation but normal reproductive capabilities were also studied. Two of the four patients were found to have serologically detectable H-Y antigen on their cells. Since H-Y antigen has been found on the cells of other patients with X chromosome abnormalities but without a Y chromosome, it is thought that the X chromosome plays a role in the regulation of H-Y antigen expression. This study suggests that the long arm of the X chromosome may be involved but the location of a regulatory gene cannot be identified in these studies. These cases do not permit us to implicate H-Y antigen as a cause of gonadal dysgenesis and early gonadal failure in females who have structurally abnormal X chromosomes.     

Fetal H-Y Typing Using Human Amniotic Fluid*

ABSTRACT: The H-Y (male) antigen is phylogenetically conserved among vertebrate species, including the species man. Previous studies have indicated the presence of a “soluble” H-Y antigen in male serum and culture fluids of male cells. We examined over 50 samples of amniotic fluid from male and female fetuses to determine if H-Y typing could be correlated with the sex of the fetus. Samples of amniotic fluid were tested to inhibit the reactivity of monoclonal antibodies in a standard H-Y assay with protein A sheep red blood cells. We found that amniotic fluids from male fetuses inhibited 40% of the reactivity and that amniotic fluids from female fetuses inhibited 0.5% of reactivity. We could also correctly identify the sex of 90% of male fetuses and 100% of female fetuses. We have not yet identified the exact nature of the inhibiting antigen(s) in the amniotic fluids, but our results clearly indicate the feasibility of fetal H-Y typing.     

Daudi supernatant, unlike other H-Y antigen sources, exerts a sex-reversing effect on embryonic chick gonad differentiation

In vitro cultures of intact chick gonads (organ cultures) and reaggregation cultures of dispersed gonad cells (roller cultures) were made. Gonads or gonad cells from 7-day-old chick embryos, at the stage when sex-specific differentiation begins, were cultured in the presence of presumed H-Y antigen-containing supernatants, or co-cultured in the presence of H-Y antigen-producing cell lines. The H-Y antigen-producing cells tested were of human, mouse, bovine and chicken origin. During organ culture, addition of supernatant of the human lymphoma cell line Daudi, or co-culture with Daudi cells, stimulated a clear proliferation of the germinal epithelium in male gonads, indicating feminization. A similar effect was obtained by treatment with estradiol. In reaggregation culture, the increase in nuclear size of germ cells was chosen as a parameter for feminization. A significant increase of germ cell nuclear size was observed in gonads cultured in the presence of Daudi supernatant. In both organ cultures and reaggregation cultures, other tested H-Y antigen sources and semi-purified H-Y antigen fractions did not exert significant effects on differentiation of the gonads or on the average area of the germ cell nuclei. These findings suggest that it is not H-Y antigen, but another protein produced by Daudi cells, that might be responsible for the sex-reversing effects.     

T cell tolerance and activation to a transgene-encoded tumor antigen

Much has been learned in recent years concerning the nature of tumor antigens recognized by T cells. To apply this knowledge clinically, the nature of the host response to individual and multiple tumor antigens has to be characterized. This will help to define the efficacy of immune surveillance and the immune status of the host following exposure to tumor antigens expressed on pre-neoplastic tissue. To approach these questions, we have developed a transgenic mouse which expresses the tumor-specific antigen P91A. The single amino acid substitution in P91A results in the expression of a new MHC class I (H-2L^d)-binding peptide. In transgenic tissue, the H-2L^d/P91A complex is expressed in isolation from other tumor-associated antigens, allowing definition of the immune response to a single defined tumor antigen, a situation closely analogous to events during tumorigenesis. We show that CD8⁺ T cell immune surveillance of P91A is ineffective without the introduction of a helper determinant operating through stimulation of CD4⁺ T cells. Recognition of the isolated P91A tumor antigen on normal tissue by CD8⁺ T cells is a tolerogenic process. Induction of T cell tolerance suggests tumor antigen-T cell interactions occurring during tumorigenesis may elicit T cell tolerance and hence confound some immunotherapeutic approaches.     

Maternal Tolerance to H-Y is Independent of IL-10

It has been suggested that the maternal immune system favors noncytotoxic, “TH-2” immune responses in order to tolerate the developing fetus. In some strains of mice, pregnant females will reject a male skin graft, even as they tolerate their male fetuses. This rejection is based on responsiveness to the male antigen H-Y. In this study we test whether functional maternal tolerance of male fetuses is critically dependent on the TH-2 cytokine Interleukin 10-(IL-10). Normal and IL-10 deficient (10-KO) females were sensitized against H-Y by intraperitoneal injection of male spleen cells before mating with 10-KO males. Litters born to 10-KO females were of comparable size to those born to normal females of the same genetic background. The proportion of males per litter was not adversely affected by IL-10 deficiency. Taken together, our work and others suggest that IL-10 may not be critically important for maternal tolerance of the fetus and extends the evidence against the idea that successful mouse pregnancy depends on TH-2 deviation of the maternal immune system.     

Major histocompatibility complex class I allele-specific peptide libraries: Identification of peptides that mimic an H-Y T cell epitope

We describe a novel method for screening large libraries of random peptides for T cell antigens. Two libraries were constructed, containing fixed amino acids representing the major histocompatibility complex (MHC) class I anchor residues for H-2K^b-restricted octamers and H-2D^b-restricted nonamers. Peptides from the K^b-restricted library (K^bL: SXIXFXXL) and the D^b-restricted library (D^bL: XXXXNXXX^I_M) specifically stabilize empty K^b and D^b molecules, respectively. The libraries contain peptides that mimic several H-2^b-restricted cytotoxic T lymphocyte epitopes, and 21 mimotopes for a D^b-restricted H-Y epitope were isolated. A degenerate synthetic peptide of limited complexity containing the identified H-Y sequence motif was found to be similar to the natural H-Y epitope by reverse-phase high performance liquid chromatography analysis. This peptide is also capable of immunizing female mice against male splenocytes. Several applications for MHC-restricted peptide libraries are discussed.     

The H-Y Response in Mid-Gestation and Long After Delivery in Mice Primed Before Pregnancy

The mechanisms underlying maternal tolerance of the semi-allogeneic fetus are not completely understood. The maternal immune system's response to the male antigen, H-Y is an example of the conflicting evidence that both supports and refutes the idea that the immune system in pregnant females is fundamentally different from that in non-pregnant females. Although multiple pregnancies may inactivate H-Y specific T cells, the immune system of the pregnant female can also generate a cytotoxic response to this antigen. To help understand this apparent conflict, we immunized female mice against H-Y with male spleen cells before pregnancy and examined the subsequent anti H-Y response during mid-pregnancy. The pregnant mice studied were able to mount cytotoxic immune responses to H-Y that were equivalent to those generated in their non-pregnant counterparts. Moreover the experience of pregnancy did not impair the ability to maintain immunologic memory to H-Y. The data support the idea that pregnancy does not violate general rules of antigen specific immunity, even if the antigen is expressed on the fetus.     

CD4+8− help prevents rapid deletion of CD8+ cells after a transient response to antigen

We have followed the fate of mature CD8⁺ T cells with a male-specific transgenic T cell receptor after antigenic stimulation with hemopoietic cells in the absence or presence of help. Our data show that mature CD8⁺ T cells can be deleted after a 3-week period of transient activation and that help, e.g. in the form of interleukin-2, can considerably delay the deletion. These experiments have implications for the design of protocols aiming at the establishment of specific immunological tolerance in T cells.     

Genetic heterogeneity of XY gonadal dysgenesis (Swyer syndrome): H-Y antigen-negative XY gonadal dysgenesis associated with inflammatory bowel disease

A 16 1/2-year-old girl was studied because of ileitis, lack of pubertal development, and primary amenorrhea. She had a 46,XY chromosome constitution in lymphocytes in fibroblasts without structural defects of X or Y. She was H-Y antigen negative. This observation supports the concept of causal heterogeneity of XY gonadal dysgenesis (Swyer syndrome). Two groups have been established: (1) H-Y antigen-positive forms, which are more common, possibly due to gonad-specific receptor defects (total failure or reduced receptor affinity), (2) H-Y antigen-negative forms possibly due to mutation in the H-Y generating system, either of the structural gene (presumably autosomal) or of a controlling gene (on the sex chromosomes). The H-Y antigen status may be of value in determining which patients are at risk for gonadoblastoma or dysgerminoma.