Dombrock genotyping in Brazilian blood donors reveals different regional frequencies of the HY allele

Background: Dombrock blood group system genotyping has revealed various rearrangements of the Dombrock gene and identified new variant alleles in Brazil (i.e., DO*A-SH, DO*A-WL and DO*B-WL). Because of the high heterogeneity of the Brazilian population, interregional differences are expected during the investigation of Dombrock genotypes. Objective: The present study aims to determine the frequencies of Dombrock genotypes in blood donors from Minas Gerais and compare the frequencies of the HY and JO alleles to those of another population in Brazil. Methods: The frequencies of the DO alleles in Minas Gerais, a southeastern state of Brazil, were determined from the genotyping of 270 blood donors. Genotyping involved polymerase chain reaction and restriction fragment length polymorphism analysis to identify the 323G>T, 350C>T, 793A>G, and 898C>G mutations, which are related to the HY, JO, DO*A/DO*B, and DO*A-WL/DO*B-WL alleles, respectively. Moreover, the frequencies of rare HY and JO alleles were statistically compared using the chi-square test with data from another Brazilian region. Results: The HY allele frequency in Minas Gerais (2.4%) was almost twice that of the JO allele (1.5%). The frequency of the HY allele was significantly higher (p-value = 0.001) than that in another Brazilian population and includes a rare homozygous donor with the Hy- phenotype. In addition, the DO*A-WL and DO*B-WL alleles, which were first identified in Brazil, were found in the state of Minas Gerais. Conclusions: The data confirm that the frequencies of DO alleles differ between regions in Brazil. The population of Minas Gerais could be targeted in a screening strategy to identify the Hy- phenotype in order to develop a rare blood bank. .

Pregnancy estrogen drives the changes of T-lymphocyte subsets and cytokines and prolongs the survival of H-Y skin graft in murine model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Estrogen as well as CD4(+)Foxp3(+) regulatory T cells were shown to have a protective role not only in maintaining maternal-fetal tolerance but also against autoimmune diseases. We aimed to investigate whether the pregnancy levels of estrogen are enough to induce transplant tolerance as to maintain fetal-maternal tolerance.</p><p><b>METHODS</b>We established H-Y skin graft transplantation in C57BL/6 ovariectomized mice that reconstituted with estrogen. Subsequently, consecutive daily estrogen injection was administrated. Tregs and the cytokines in the peripheral blood were detected by flow cytometry and ELISA pre- and post-transplant.</p><p><b>RESULTS</b>The results indicated that pregnancy levels of estrogen could promote Tregs in secondary lymphoid organs and peripheral blood (P < 0.05) but not thymus (P > 0.05). The estrogen-treated recipients accepted H-Y skin grafts for more than 35 days (median survival time (MST): (44.0 ± 1.2) days) compared with estrogen-untreated mice (MST: (23.0 ± 1.6) days) (P < 0.05). It was also observed that estrogen up-regulated the expression of Foxp3, but did not affect CD3(+)CD8(+) effector T-cells in non-transplant mice. While in the presence of H-Y antigens, the expression of Foxp3 was more significant and CD3(+)CD8(+) effector T cells were decreased significantly (P < 0.05). Meanwhile, the up-regulated IL-10 and IL-4, and down-regulated IFN-γ could be observed (P < 0.05).</p><p><b>CONCLUSIONS</b>Pregnancy levels of estrogen may promote the conversion of peripheral Tregs in secondary lymphoid organs, but show no effect on the natural Tregs production, differentiation and maturity in central lymphoid organs. Furthermore, pregnancy levels of estrogen could significantly prolong the survivals of H-Y skin grafts by the expansion of Tregs, suppression of CD3(+)CD8(+) effector T-cells and immune shift towards Th2 cytokines.</p>

Falha na sexagem por inibição do desenvolvimento de embriões bovinos produzidos in vitro com anticorpos anti H-Y / Failure of sexing by developmental arrest of bovine embryos in vitro produced with H-Y antisera

Embriões bovinos produzidos in vitro, em estádio de mórula, foram cultivados em meio contendo anticorpos anti H-Y de alto título proveniente de ratos por 24h e, após este tempo, classificados em dois grupos: 1) embriões inibidos em estádio de mórula (classificados como machos) e 2) embriões que se desenvolveram e formaram a blastocele (classificados como fêmeas). O sexo de 311 embriões, distribuídos em três grupos de concentração dos anticorpos, 3 por cento, 5 por cento ou 7 por cento, foi identificado pela reação em cadeia da polimerase. Não houve desvio da proporção entre machos e fêmeas (P>0,05) nos grupos em que se utilizaram os anticorpos anti H-Y, quando comparadas ao grupo-controle, sem adição de anticorpos anti H-Y. Diferentemente dos resultados obtidos utilizando-se embriões bovinos produzidos in vivo, a sexagem com anticorpos anti H-Y de alto título em embriões produzidos in vitro não propiciou sucesso.

In vitro produced bovine embryos at morula stage were cultured in medium containing high titer of rat H-Y antisera for 24h. The embryos were classified in two groups: 1) embryos arrested at morula stage (classified as males); and 2) embryos that developed and formed a blastocoele (classified as female). The sex of 311 embryos, divided in three groups of concentration of H-Y antisera, 3 percent, 5 percent or 7 percent, was identified by polimerase chain reaction. The results showed no difference (P>0.05) on sexual deviation in groups in which the H-Y antisera was added, in relation to control group, in which no H-Y antisera was added. In contrast with results obtained with in vivo produced bovine embryos, the sexing of in vitro produced bovine embryos with high H-Y antisera titer did not succed.

Cloning and analysis of phage Fab antibodies of mouse male specific antigen / 生物工程学报

To clone mouse phage antibodies against H-Y antigen from a phage antibody library, three cycles of affinity enrichment of the mouse phage antibody library with male spleen cells and two cycles of nonspecific absorption with female spleen cells were performed. The presence of mouse Fab on the phage surface was determined by ELISA and sequence analysis. 9 of 15 strains can bind to male spleen cells with the specific activity. Recombination rate of the phage antibody library clones is 60%. Sequence analysis of the PCR products of plasmid DNA of E5 clones show VH and Vkappa had common characteristics shared by other known variable region of antibodies. The mouse phage Fab antibody could be used for identifying H-Y antigen, and for the development of sex determination of early embryos in mammals.

Immune Tolerance in Murine Islet Transplantation Across HY Disparity

BACKGROUND: Minor histocompatibility HY antigen, as a transplantation antigen, has been known to cause graft rejection in MHC (major histocompatibility complex) matched donor-recipient. The aim of our study is to investigate the role of male antigen (HY) disparity on MHC matched pancreatic islet transplantation and to examine the mechanism of the immune reaction. METHODS: Pancreatic islets were isolated and purified by collagen digestion followed by Ficoll gradient. The isolated islets of male C57BL6/J were transplanted underneath the kidney capsule of syngeneic female mice rendered diabetic with streptozotocine. Blood glucose was monitored for the rejection of engrafted islets. After certain period of time, tail to flank skin transplantation was performed either on mouse transplanted with HY mismatched islets or on sham treated mouse. The rejection was monitored by scoring gross pathology of the engrafted skin. RESULTS: HY mismatched islets survived more than 300 days in 14 out of 15 mice. The acceptance of second party graft (male B6 islets) and the rejection of third party graft (male BALB/c islets) in these mice suggested the tolerance to islets with HY disparity. B6 Skin with HY disparity was rejected on day 25 +/- 7. However, HY mismatched skin transplanted on the mice tolerated to HY mismatched islets survived more than 240 days. Tetramer staining in these mice indicated the CTL recognizing MHC Db/Uty was not deleted or anergized. CONCLUSION: The islet transplantation across HY disparity induced tolerance to HY antigen in C57BL6 mouse, which in turn induced tolerance to HY mismatched skin, which otherwise would be rejected within 25 days. The MHC tetramer staining suggested the underlying mechanisms would not be clonal deletion or anergy.

FEMALE PSEUDOHERMAPHRODITISM DUE TO ADRENOGENITAL SYNDROME

Female pseudohermaphroditism due to adrenogenital syndrome is a condition in which individuals with a 46XX karyotype, negative H-Y antigen, normal mullerian duct derivatives, and a lack of development of w lffian duct structures differentiate partially as phenotypic males. They usually manifest masculinization of the external genitalia as a result of excess endogenous androgens. Most female pseudohermaphrodities have one of the types of congenital virilizing adrenal hyperplasia. Adrenogenital syndrome is inborn errors transmitted by autosomal recessive genes and may be due to defects in any of the enzymic steps in the biosynthesis of cortisol. Most affected individuals have a failure of 21-hydroxylation which prevents the conversion of 17 alpha-hydroxyprogesterone to 11-deoxycortisol. Such a defect in 21-hydroxylase leads to excessive production of adrenal androgens causing virilization. The treatment is early endocrinologic support and surgical reconstruction. There are some considerations in surgical repairs including normal sized clitoris with adequate erogenous sensation, sufficiently wide vaginal introitus and normal aesthetic appearance of the external genitalia for her normalized life as a female. We have experienced four cases of female pseudohermaphroditism due to adrenogenital syndrome. In two cases, we performed clitoroplasty with nerve sparing technique, vulvoplasty with mons pubis augmentation, vaginoplasty with posterior perineal flap and urethral reconstruction. In the other cases, we performed clitoroplasty with nerve sparing technique, vulvoplasty and vaginoplasty There was no specific operative complication and the result of the correction was satisfactory.

Papel do cromossomo Y na diferenciaçäo gonadal / Y chromosome role in the gonadal differentiation

Os autores fazem uma revisao sobre a evoluçao dos conceitos de diferenciaçao gonadal, desde a descoberta dos cromossomos X e Y até a localizaçao da seqüência de bases, no braço curto do cromossomo Y, próximo à regiao pseudoautossômica, supostamente responsável pela diferenciaçao da gônada bipotencial a testículo. O assunto desperta grande interesse, pois nem todos os passos deste verdadeiro quebra-cabeças foram desvendados e a questao do virilismo XX ainda se acha aberta à pesquisa científica.

A case of XX male syndrome / 대한비뇨기과학회지

The 46, XX male or sex-reversal syndrome is a rare entity, which may be reported first by de la Chapelle and associates in 1964, an additional 135 cases have been recognized, yet only 20 percent of these patients have been diagnosed during childhood. The 46, XX male may be associated with hypogonadism and infertility in adult, and occasionally, sexual ambiguity in the neonate. At least 10% of patients have had hypospadia or ambiguous external genitalia. The 46, XX male was diagnosed with cytogenic study, H-Y antigen, hormonal study testicular biopsy, radiologic study. Here, we report a case of 19 month-old child XX-male with hypospadia and chordee.

Soluble H-Y antigen and the search for molecules controlling sex determination in mammals

Recentemente demonstrou-se que a linhagem celular TM-4, derivada de células de Sertoli isoladas do testículo de camundongos imaturos BALB/c, secreta o antígeno H-Y. Essa evidência vem em apoio à hipótese de que o antígeno H-Y é o indutor do testículo em mamíferos. A disponibilidade de uma fonte de antígeno H-Y solúvel permitiu-nos desenvolver um imunoensaio sensível para a determinaçäo do antígeno H-Y em pacientes intersexuados. No entanto, a pesquisa de moléculas que controlam a determinaçäo do sexo em mamíferos (e em outros vertebrados) permanece um campo controvertido. O uso de sondas moleculares para o cromossomo Y, em conjunto com testes sorológicos, poderá permitir evidências mais conclusivas num futuro próximo

Digressöes teleológicas bioplásticas / Bioplastical teleological digressions

O autor passa em revista uma série de fatos interessantes que pôde observar através de uma longa vivência no âmbito da cirurgia plástica. Procura interpretar fenômenos curiosos e muitas vezes paradoxais que acompanham a evoluçäo dos transplantes cutâneos, ósseos, corneanos, endócrinos e outros. Procura mostrar a importância da especificidade genética dos transplantes e o seu comportamento correlato, bem como as ilaçöes que delas podem ser tiradas. Aborda, a seguir, sucintamente o fenômeno da "tolerância" aos enxertos, o fenômeno da chamada "doença imunológica" e aventa uma hipótese de trabalho concernente ao antígeno H-Y

Estudo clinico, citogenetico, histologico e da regulacao hormonal em dois casos de hermafroditismo verdadeiro. / Clinical, cytogenetic, hystological and hormonal regulation studies in 2 cases of true hermaphroditism

Foi realizado estudo clinico, citogenetico histologico e da secrecao hormonal em dois casos de hermafroditismo verdadeiro (HV).O primeiro paciente apresentava cariotipo XX e o segundo XX/XY, sendo os dois antigeno HY positivo. O exame histologico revelou ovotestis no primeiro caso com presenca de espermatogonias ate agora nao descritas no HV. O segundo paciente apresentava ovario histologicamente normal e testiculo com areas de hialinizacao tubular. A avaliacao hormonal no paciente 1 revelou grandes flutuacoes dos niveis de LH, FSH, estradiol e progesterona. Os dois pacientes apresentavam baixa resposta testicular ao estimulo com gonadotrofina corionica e liberacao de LH apos uso de estrogeno semelhante a observada no ser feminino.O citrato de clomifeno induziu resposta ovulatoria no caso testado, podendo ser utilizado na identificacao da presenca de ovotestis em caso de suspeita de HV

O antigeno H-Y e a diferenciacao sexual do cerebro. / The H-Y antigen and the sex differentiation of the brain

Informacoes recentes sobre a expressao do antigeno H-Y em transexuais tem demonstrado que a maioria desses pacientes possui a expressao do antigeno H-Y compativel com a do sexo ao qual eles desejam pertencer. E portanto provavel que o antigeno H-Y desempenhe um papel na determinacao da identidade sexual, em adicao a sua reconhecida acao na determinacao do sexo gonadal.Neste artigo nos propomos um modelo para a acao do antigeno H-Y na diferenciacao sexual do cerebro