RESUMO
We present the case of a Dutch family with a new mutation (c523_528dup) in GATA3 causing HDR syndrome. HDR syndrome is characterised by hypoparathyroidism, deafness and renal defects. In this study, we describe the audiometric characteristics of 5 patients from this family. Their hearing impairment was congenital, bilateral and symmetric. Audiograms showed mild-to-moderate hearing impairment with a flat audiogram configuration. Higher frequencies tended to be affected more strongly. Cross-sectional analyses showed no progression, and a mean audiogram was established. Psychophysical measurements in 3 HDR patients - including speech reception in noise, loudness scaling, gap detection and difference limen for frequency - were obtained to assess hearing function in greater detail. Overall, the results of the psychophysical measurements indicated characteristics of outer hair cell loss. CT scanning showed no anomalies in 3 of the HDR patients. Although 2 patients displayed vestibular symptoms, no anomalies in the vestibular system were found by vestibulo-ocular examination. Our results are in agreement with the theory that outer hair cell malfunctioning can play a major role in HDR syndrome.
Assuntos
Fator de Transcrição GATA3/genética , Perda Auditiva Neurossensorial/genética , Hipoparatireoidismo/genética , Mutação , Nefrose/genética , Audiometria de Tons Puros , Feminino , Perda Auditiva Neurossensorial/fisiopatologia , Humanos , Hipoparatireoidismo/fisiopatologia , Masculino , Nefrose/fisiopatologia , Países Baixos , Linhagem , Fenótipo , Percepção da Fala/fisiologia , Síndrome , Testes de Função VestibularRESUMO
BACKGROUND: Seminal plasma contains signaling molecules capable of modulating the maternal immune environment to support implantation and pregnancy. Prior studies indicated that seminal plasma induces changes in gene transcription of maternal immune cells. Reduced immune suppressive capacity may lead to pregnancy loss. The aim of this study was to investigate the immunomodulating effects of seminal plasma on T cells and monocytes in the context of recurrent pregnancy loss (RPL). METHODS: Female T cells and monocytes were incubated with seminal plasma of 20 males in unexplained RPL couples (RPL males) and of 11 males whose partners had ongoing pregnancies (control males). The effect of seminal plasma on messenger RNA (mRNA) expression of immune cells was measured. Levels of mRNA expression were related to key signaling molecules present in the seminal plasma. Agglomerative hierarchical cluster analysis was performed on seminal plasma expression profiles and on mRNA expression profiles. RESULTS: Expression of CD25 and anti-inflammatory IL-10 by female T cells was significantly lower after stimulation with seminal plasma of RPL males compared to control males. Female monocytes treated with seminal plasma of RPL males showed an immune activation signature of relatively elevated HLA-DR expression. Expression of these T cell and monocyte components was particularly correlated with the amounts of TGF-ß and VEGF in the seminal plasma. CONCLUSION: Our findings indicate that seminal plasma has immunomodulating properties on female immune cells compatible with the induction of a more regulatory phenotype, which may be impaired in cases of unexplained RPL.
Assuntos
Aborto Habitual , Sêmen , Feminino , Humanos , Imunomodulação , Masculino , Gravidez , RNA Mensageiro/metabolismo , Linfócitos T/metabolismoRESUMO
BACKGROUND: Seminal plasma contains a wide range of cytokines, chemokines and growth factors. Part of these signalling molecules assist in inducing a state of active maternal immune tolerance towards the fetus. Disbalances in seminal plasma content may contribute to pregnancy loss. This study investigated cytokine expression profiles in seminal plasma of male partners of couples with unexplained recurrent pregnancy loss (RPL) and the association with clinical and lifestyle characteristics, including smoking, alcohol consumption and body mass index (BMI). METHODS: In the seminal plasma of 52 men who visited a specialised RPL clinic the levels of 25 pre-selected cytokines, chemokines and growth factors were measured by Bio-Plex assay or ELISA. Two-way hierarchical cluster analysis was performed. Identified patient clusters were compared on clinical and lifestyle characteristics. RESULTS: Two distinct cytokine expression profiles in the seminal plasma were revealed by cluster analysis. Patient cluster I showed relatively higher levels of pro-inflammatory cytokines, including IL-1α, IL-1ß, IL-6, IL-8, IL-12, IL-18 and TNF-α, compared to Patient cluster II. Men belonging to Patient cluster I were significantly older and had significantly more lifestyle risk factors compared to men in Patient cluster II. CONCLUSION: Cluster analysis suggested the existence of a less favourable pro-inflammatory cytokine expression profile, being present in part of men affected by RPL and associated with advanced male age and lifestyle risk factors. These findings may serve as a starting point for further research into underlying mechanisms and ultimately lead to novel diagnostic and therapeutic approaches for couples with RPL.
Assuntos
Aborto Habitual/diagnóstico , Citocinas/análise , Sêmen/imunologia , Aborto Habitual/imunologia , Adulto , Fatores Etários , Biomarcadores/análise , Biomarcadores/metabolismo , Estudos de Casos e Controles , Citocinas/metabolismo , Feminino , Voluntários Saudáveis , Humanos , Masculino , Gravidez , Prognóstico , Sêmen/metabolismo , Análise do Sêmen/métodosRESUMO
OBJECTIVE: Investigating the association between plasma levels of cytokines and chemokines, Selenoprotein S (SELS) gene variation and osteoarthritis (OA) subtypes. METHODS: The genetics of osteoarthritis and progression (GARP) study consists of 191 sibling pairs with symptomatic OA at multiple joint sites. We have measured plasma levels of 17 cytokines and chemokines and genetic variation at the SELS gene. RESULTS: Nine out of 17 serum markers could be assessed quantitatively, whereas eight markers were assessed qualitatively. Principal component analysis (PCA) on the quantitatively assessed markers and serum high sensitive C-reactive protein (S-HsCRP) revealed that three components underlie 61% of the total plasma variation. Three single nucleotide polymorphisms (SNPs) in the SELS gene revealed four common haplotypes, one of which, GAG (frequency 3.5%) showed significant association to an anti-inflammatory (P=0.019) and acute phase related (P=0.036) component. OA subtype analysis showed that one component (mainly representing chemokine variation) was significantly associated to hand OA and disc degeneration (P=0.029 and P=0.010 respectively) as well as a physical component score (PCS) (P=0.042). The CRP related component also showed a strong association to the PCS (P=0.007). SELS haplotypes showed no association to OA subtypes in the GARP study. CONCLUSION: Genetic variation in the SELS gene associates to components representing inflammatory signaling. Another component, representing chemokine variation, showed association to hand OA and disc degeneration in the GARP study indicating chemokines may contribute to OA pathogenesis.
Assuntos
Proteína C-Reativa/metabolismo , Citocinas/sangue , Osteoartrite/sangue , Selenoproteínas/metabolismo , Líquido Sinovial/metabolismo , Adulto , Idoso , Biomarcadores/metabolismo , Proteína C-Reativa/genética , Citocinas/genética , Feminino , Variação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite/genética , Selenoproteínas/genética , IrmãosRESUMO
PROBLEM: The short term effect of the caesarean delivery on the phenotypic and functional characteristics of the peripheral blood leukocytes of the mother is unknown. METHOD OF STUDY: We determined the composition and activation status of the maternal peripheral blood leukocytes isolated within 4h before and within 24h after elective caesarean delivery with neuraxial anaesthesia. Furthermore, we determined the proliferative and cytotoxic response of these leukocytes to several stimulators. RESULTS: No significant differences in the percentage of CD4+CD25bright and CD8+CD28- T cells or the expression of activation markers FoxP3, CD69 and HLA-DR were observed in peripheral blood drawn before caesarean delivery compared to after caesarean delivery. Also the alloreactive immune responses in samples taken before and after the caesarean delivery were similar. CONCLUSION: Our results show that the phenotype and immune response of maternal peripheral blood T cells obtained before elective caesarean delivery are not different from those obtained after caesarean delivery. This knowledge will facilitate sample collection for future studies on the immune response in term pregnancies.
Assuntos
Células Sanguíneas/imunologia , Cesárea , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Proliferação de Células , Citotoxicidade Imunológica , Feminino , Fatores de Transcrição Forkhead/metabolismo , Humanos , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Isoantígenos/imunologia , Ativação Linfocitária , Mães , GravidezRESUMO
HLA-C is the only polymorphic classical HLA I antigen expressed on trophoblast cells. It is known that higher incidence of C4d deposition on trophoblast cells is present in women with recurrent miscarriage. C4d is a footprint of antibody-mediated classical complement activation. Therefore, this study hypothesize that antibodies against HLA-C may play a role in the occurrence of unexplained consecutive recurrent miscarriage. Present case control study compared the incidence of HLA-C specific antibodies in 95 women with at least three consecutive miscarriages and 105 women with uneventful pregnancy. In the first trimester of the next pregnancy, presence and specificity of HLA antibodies were determined and their complement fixing ability. The incidence of HLA antibodies was compared with uni- and multivariate logistic regression models adjusting for possible confounders. Although in general a higher incidence of HLA antibodies was found in women with recurrent miscarriage 31.6% vs. in control subjects 9.5% (adjusted OR 4.3, 95% CI 2.0-9.5), the contribution of antibodies against HLA-C was significantly higher in women with recurrent miscarriage (9.5%) compared to women with uneventful pregnancy (1%) (adjusted OR 11.0, 95% CI 1.3-89.0). In contrast to the control group, HLA-C antibodies in the recurrent miscarriage group were more often able to bind complement. The higher incidence of antibodies specific for HLA-C in women with recurrent miscarriage suggests that HLA-C antibodies may be involved in the aetiology of unexplained consecutive recurrent miscarriage.
Assuntos
Aborto Habitual/imunologia , Antígenos HLA-C/metabolismo , Trofoblastos/metabolismo , Adulto , Anticorpos/metabolismo , Citotoxicidade Celular Dependente de Anticorpos , Estudos de Casos e Controles , Ativação do Complemento , Complemento C4b/metabolismo , Feminino , Antígenos HLA-C/imunologia , Humanos , Fragmentos de Peptídeos/metabolismo , Gravidez , Primeiro Trimestre da Gravidez , Ligação ProteicaRESUMO
The maternal immune system must adapt to tolerate the invasion of the allogeneic feto-placental unit. It is generally accepted that improper adaptation causes pregnancy complications like preeclampsia. The Epstein-Barr virus-induced gene 3 (EBI3) protein is a subunit of immune-modulatory cytokines interleukin 27 (IL-27) and IL-35. EBI3 has been reported to associate with HLA-G. In this small pilot study we find higher decidual EBI3 (p<0.05) and HLA-G (p<0.01) mRNA expression in preeclampsia (n=7) compared to normotensive (n=8) pregnancies. Whether the higher EBI3 and HLA-G mRNA expression is a consequence or cause of preeclampsia remains to be answered. Further research to determine the effects on IL-27 and IL-35 is needed.
Assuntos
Decídua/metabolismo , Antígenos HLA-G/metabolismo , Interleucinas/metabolismo , Pré-Eclâmpsia/imunologia , Adulto , Feminino , Antígenos HLA-G/genética , Humanos , Interleucina-27/genética , Interleucinas/genética , Pessoa de Meia-Idade , Antígenos de Histocompatibilidade Menor , Projetos Piloto , Pré-Eclâmpsia/genética , Gravidez , Tolerância ao Transplante , Regulação para Cima , Adulto JovemRESUMO
Seminal plasma (SP) contains immunomodulatory factors that may contribute to the formation of a tolerogenic environment at the embryo implantation site. The main focus of this study was to investigate the influence of SP on female T cells in the presence and absence of antigen-presenting cells (APCs) in an in vitro model. Female PBMCs and T cells were incubated with SP from seminal fluid samples of known and variable sperm quality. The immediate effect of SP on the mRNA expression of CD25, IL-10, IFN-γ, and Foxp3 was measured. Furthermore, proliferative responses, cytokine production, and CD25 expression were determined. Exposure to SP leads to increased mRNA expression of CD25, IL-10, and Foxp3 in T cells. Induction of mRNA for IL-10 and CD25 was dependent on the presence of APCs. Both PBMCs and T cells exposed to SP showed a proliferative response and produced several cytokines. The proliferative effects of SP on T cells observed were independent of sperm quality parameters, cytokines or soluble HLA molecules in SP. Furthermore, the presence of SP induced a higher expression of CD25 on the membrane of CD4+ T cells. SP has a direct immunomodulatory effect on T cells, as reflected in a proliferative response and upregulation of Foxp3. The presence of APCs is needed to induce IL-10 and CD25 upregulation in T cells exposed to SP. In conclusion, SP has both a direct and an indirect effect mediated through APCs on T cells.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Proliferação de Células , Imunomodulação , Sêmen/imunologia , Adulto , Linfócitos T CD4-Positivos/citologia , Feminino , Antígenos HLA/imunologia , Humanos , Interleucina-10/imunologia , Subunidade alfa de Receptor de Interleucina-2/imunologia , Masculino , Regulação para Cima/imunologiaRESUMO
BACKGROUND: Acceptable HLA mismatches for highly sensitized patients are determined so as to increase their chances of receiving transplants. The disadvantage of the current procedures is that the antibody reactivity of the patients' sera is tested against HLA antigens expressed on cells or HLA antigens isolated from cell lysates. Therefore, two (homozygous for HLA-A and -B) to four (heterozygous for HLA-A and -B) different HLA class I antigens are present in the test. This might cause reactivity toward nonacceptable mismatches to mask the determination of acceptable mismatches. METHODS: Recently we observed that the detection of soluble HLA class I antigens is inhibited by HLA-specific antibodies. In the present study, inhibition of soluble HLA-specific ELISAs (anti-soluble HLA-A2, -B7, -B12) was evaluated as a tool used to determine acceptable mismatches. The results were compared with current determination of acceptable mismatches (which is by complement-dependent cytotoxicity and/or fluorescence-activated cell sorter analysis). RESULTS: In the case of acceptable mismatches determined by conventional methods, sera from the patients were not interfering in these ELISAs, whereas in the case of nonacceptable mismatches (thus specific antibodies), significant inhibition was observed in most instances. Among the nonacceptable mismatches, the test showed significant inhibition in 20 of 24 cases, whereas among acceptable mismatches, no inhibition was observed (in eight of eight), indicating the lack of specific antibodies. CONCLUSIONS: In highly sensitized patients, the introduction of soluble HLA-specific ELISAs is of additional and confirmatory value for the determination of acceptable mismatches. The major advantage of this approach is that antibody reactivity is tested against single antigens only.
Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Antígenos HLA/sangue , Antígenos de Histocompatibilidade Classe I/sangue , Anticorpos/imunologia , Especificidade de Anticorpos , Tipagem e Reações Cruzadas Sanguíneas , Teste de Histocompatibilidade , Humanos , Imunização , SolubilidadeRESUMO
In healthy individuals, like in highly sensitized patients, CTL precursors (CTLp) frequencies against HLA-A antigens were found to be significantly lower than CTLp frequencies against HLA-B antigens. In the present study, we tested whether not only quantitative but also qualitative differences exist between CTLs against HLA-A antigens versus HLA-B antigens. Experiments performed in the presence of antibodies against CD8 showed that CTLp directed against HLA-B antigens were significantly less inhibited by anti-CD8 compared with CTLp against HLA-A. The low anti-CD8 inhibition of the CTLp directed against HLA-B antigens may be explained by a higher affinity of the CTLp for HLA-B antigens on the target cells. CTLp directed against HLA-B antigens were also found to be less sensitive to CsA, which suggests that their high affinity is due to previous priming. A clinical relevance of these findings is suggested by the observation that matching for HLA-B antigens is of greater importance for kidney graft survival than matching for HLA-A.
Assuntos
Antígenos HLA-A/imunologia , Antígenos HLA-B/imunologia , Linfócitos T Citotóxicos/imunologia , Rejeição de Enxerto/prevenção & controle , Antígenos HLA-A/análise , Antígenos HLA-B/análise , Teste de Histocompatibilidade , Humanos , Transplante de Rim/imunologia , Células-Tronco/imunologiaRESUMO
In order to test whether a selective T cell nonresponsiveness to noninherited maternal human leukocyte antigens (NIM) exists, we measured the frequencies of alloreactive T cells of healthy individuals to their NIM HLA antigens and to their noninherited paternal (NIP) HLA antigens by limiting dilution assays. Both the frequencies of cytotoxic T cell precursors (CTLp) and IL-2-producing helper T cell precursors (HTLp) were determined. Similar frequencies were observed for NIM class I-reactive CTLp and NIP class I-reactive CTLp. This was the case when frequencies were determined against total NIM and NIP haplotypes but also when CTLp frequencies against individual NIM and NIP antigens were measured. A positive finding of this study was the confirmation of our earlier observation that CTLp frequencies against individual HLA-A antigens are generally lower than CTLp frequencies against HLA-B. This was found both for the maternal and the paternal HLA-A and -B antigens. Similarly, comparable frequencies of IL-2-producing helper T cell precursors directed against NIM HLA class II antigens and NIP HLA class II antigens were found. When breast-feeding in the neonatal period was considered, no differences in the frequencies of CTLp and HTLp were observed between children who had been breast-fed and children who had not. Therefore the present data do not support the hypothesis that confrontation with noninherited maternal HLA in neonatal life leads to down-regulation of alloreactive T cell responses to the mother.
Assuntos
Antígenos HLA/imunologia , Imunidade Materno-Adquirida/imunologia , Linfócitos T/imunologia , Adulto , Criança , Epitopos , Pai , Feminino , Haplótipos , Humanos , Interleucina-2/farmacologia , Masculino , Células-Tronco/efeitos dos fármacos , Células-Tronco/imunologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos , Linfócitos T Auxiliares-Indutores/imunologiaRESUMO
Although the cornea is considered to be an immunological privileged site, corneal transplantation can result in immunological rejection followed by graft failure, especially in patients with vascularized corneas. Several studies suggest a beneficial effect of matching for the HLA class I antigens on corneal graft survival, although a large study (the Collaborative Corneal Transplantation Study) failed to confirm this. To circumvent an endless discussion on studies either confirming or denying the relevance of HLA matching, we decided to approach this problem in another way. A more direct way to assess the importance of HLA class I antigens in corneal transplantation is to measure whether rejection of an allograft is associated with priming of cytotoxic T lymphocytes recognizing the mismatched HLA antigens of the donor. In the present study, 13 patients with good graft function and 10 with ongoing rejection of their corneal allografts were analyzed for the presence of CTL directed against mismatched donor HLA class I antigens, by limiting dilution assays. CTLs were divided into naive and primed CTLs based on the measurement of their in vitro sensitivity or resistance to anti-CD8 or cyclosporine. Cytotoxic T cell precursor frequencies directed against the mismatched donor HLA class I antigens were similar in nonrejectors and rejectors. However, rejection was strongly associated with the presence of primed, donor-specific CTL, whereas these primed cells were absent in case of good graft function. These data show that HLA antigens of a transplanted cornea are immunogenic and targets for rejection by cytotoxic T cells. Therefore, this study supports the need for HLA-A and -B matching in corneal transplantation in patients with a high probability of rejection.
Assuntos
Transplante de Córnea/imunologia , Rejeição de Enxerto/imunologia , Antígenos de Histocompatibilidade Classe I/análise , Linfócitos T/imunologia , Doadores de Tecidos , Anticorpos/imunologia , Formação de Anticorpos , Antígenos CD8/imunologia , Transplante de Córnea/fisiologia , Ciclosporina/farmacologia , Método Duplo-Cego , Humanos , Técnicas de Diluição do Indicador , Ativação Linfocitária/efeitos dos fármacos , Células-Tronco , Linfócitos T Citotóxicos/citologia , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/imunologiaRESUMO
Pre-existing alloantibodies against the mismatched HLA-A and HLA-B antigens of the donor, when present in current sera, are believed to be detrimental to kidney graft survival. When these antibodies are present only in historical sera, their immunoglobulin class has reported to be important with respect to the expected graft survival, IgG antibodies being associated with poor graft prognosis and IgM with reasonable graft survival. In the present study, we have tested whether the immunoglobulin class of anti-HLA antibodies is reflected in the activation state of CTLs directed against these HLA antigens as measured by their in vitro resistance or sensitivity to CsA. The results indicate that the presence of IgG anti-HLA antibodies is associated with the presence of activated CTLs (CsA resistant), whereas in the case of IgM antibodies, mainly naive CTLs (CsA sensitive) are found. This observation may explain the different prognoses of historical positive crossmatches due to IgG versus IgM alloantibodies.
Assuntos
Antígenos de Histocompatibilidade Classe I/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Linfócitos T Citotóxicos/imunologia , Ciclosporina/farmacologia , Citotoxicidade Imunológica/efeitos dos fármacos , Humanos , Isoanticorpos/imunologia , Transplante de Rim/imunologia , Linfócitos T Citotóxicos/efeitos dos fármacosRESUMO
PURPOSE: To identify the presence of membrane-bound regulators of complement activation (m-RCA) on uveal melanomas and uveal melanoma cell lines and to examine their role in the inhibition of complement-mediated lysis in vitro. METHODS: Immunohistochemistry and flow cytometric analysis with monoclonal antibodies directed against m-RCA CD46, CD55, and CD59 were applied to tissue sections of 10 uveal melanomas, three primary uveal melanoma cell lines, and one uveal melanoma metastatic cell line. A microcytotoxicity test was used for measuring antibody-dependent complement-mediated lysis. RESULTS: The tissue sections and all four uveal melanoma cell lines expressed CD46, CD55, and CD59. Complement-mediated lysis in the presence of human complement was increased after partial removal of the m-RCA CD55 and CD59 with phosphatidylinositol-specific phospholipase C from the uveal melanoma cell line 92-1. CONCLUSIONS: These results demonstrate that CD46, CD55, and CD59 are expressed in uveal melanomas and that CD55 or CD59, or both, plays a role in resistance to complement-mediated cytotoxicity. The finding that m-RCA are expressed in uveal melanomas may have implications for the effectiveness of the anti-tumor response and in the therapeutic application of monoclonal antibodies directed against tumor-associated antigens.
Assuntos
Antígenos CD/análise , Antígenos CD55/análise , Antígenos CD59/análise , Ativação do Complemento , Proteínas Inativadoras do Complemento/análise , Melanoma/imunologia , Glicoproteínas de Membrana/análise , Neoplasias Uveais/imunologia , Idoso , Anticorpos Monoclonais , Citotoxicidade Celular Dependente de Anticorpos , Linhagem Celular , Membrana Celular/imunologia , Membrana Celular/patologia , Neoplasias da Coroide/imunologia , Neoplasias da Coroide/patologia , Feminino , Citometria de Fluxo , Antígenos de Histocompatibilidade Classe I/análise , Humanos , Imuno-Histoquímica , Linfócitos/imunologia , Masculino , Melanoma/patologia , Proteína Cofatora de Membrana , Pessoa de Meia-Idade , Neoplasias Uveais/patologiaRESUMO
PURPOSE: To examine the role of donor-specific antibodies, with or without complement, in rejection of orthotopic corneal transplants by using mice as recipients in which the genes for the heavy chain of immunoglobulin or the third complement component have been eliminated by homologous recombination. METHODS: BALB/c corneas were transplanted into eyes of B-cell-deficient (n=17) or wild-type control C57BL/6 (n=30) mice and into eyes of complement (C3)-deficient (n=15) or wild-type control 129-C57BL/6 (n=13) mice. After surgery all grafts were evaluated over 8 weeks in a masked manner by biomicroscopy for signs of rejection. RESULTS: The rates of corneal transplant rejection were similar among B-cell-deficient and C3-deficient mice compared with rejection rates in their respective wild-type control subjects. This similarity applied to the time course of rejection and to cumulative survival rates. CONCLUSIONS: Neither donor-specific antibody nor the third component of complement play essential roles in acute rejection of orthotopic corneal allografts in mice.
Assuntos
Linfócitos B/fisiologia , Transplante de Córnea , Genes de Imunoglobulinas , Cadeias Pesadas de Imunoglobulinas/genética , Síndromes de Imunodeficiência/genética , Animais , Complemento C3/deficiência , Complemento C3/fisiologia , Transplante de Córnea/imunologia , Genes de Imunoglobulinas/fisiologia , Rejeição de Enxerto/imunologia , Sobrevivência de Enxerto/fisiologia , Síndromes de Imunodeficiência/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BLRESUMO
Previous studies showed that pregnancy can prime the maternal cellular immune response directed against paternal HLA antigens. Primed CTLs specific for inherited paternal HLA antigens (IPA) were found in women who had formed HLA allo antibodies, whereas naive CTLs were present in women who did not form antibodies against the paternal HLA antigens. As HLA allo antibodies may disappear in time, it is not clear which women on the waiting list for transplantation have been sensitized to paternal HLA antigens and are at risk for graft rejection if paternal HLA antigens are shared by the donor organ. The presence of primed CTLs specific for a particular antigen is considered to be a reflection of sensitization.In the present study we investigated whether these primed CTLs persist in women who had been pregnant and had formed antibodies against the inherited paternal HLA class I antigens. For this purpose 14 women who had their last pregnancy 10 years ago were analyzed with respect to IPA-specific CTLp frequencies and the presence of high avidity CTLs directed against inherited paternal HLA class I antigens. Although primed CTLs specific for IPA's were found more frequently in women with persisting alloantibodies, they still can be detected when the antibodies have disappeared. The current data show that primed CTLs directed against inherited paternal HLA antigens towards which antibodies have been formed in the past can persist for more than 10 years after pregnancy. The cellular test used in our study can be useful to detect presensitization in women with a history of pregnancy, who enter the waiting list for transplantation.
Assuntos
Antígenos HLA/imunologia , Linfócitos T Citotóxicos/imunologia , Feminino , Humanos , Masculino , Gravidez , Fatores de TempoRESUMO
The introduction of molecular tissue typing techniques has led to an enormous increase in the number of human leucocyte antigen (HLA) alleles. This increasing polymorphism of the HLA antigens makes the selection of a well-matched unrelated donor a difficult task. Recent data suggest that some HLA mismatches are more immunogenic than others. This has led to the introduction of terms like beneficial or acceptable versus detrimental or taboo mismatches. The study considered whether the differential immunogenicity as reflected by graft survival studies can be detected in vitro as well. Mixed lymphocyte reaction (MLR) and primed lymphocyte tests (PLT) were performed with different HLA-DR mismatched combinations in the presence and absence of cyclosporine A and prednisolone. Differential effects of these immunosuppressive drugs were observed. Some reactions could easily be blocked by cyclosporine alone, whereas others need the addition of high doses of prednisolone as well before a significant inhibition was found. These differences were not only found between individual responders but also within one individual dependent on the stimulatory HLA-antigen involved. When the group of beneficial mismatches was compared with the group of detrimental mismatches, no differences were observed. Our data show that immunosuppressive drugs have a differential effect on in vitro alloimmune responses but these do not differentiate between beneficial and detrimental mismatches as defined by kidney graft survival.
Assuntos
Ciclosporina/farmacologia , Antígenos HLA-DR/imunologia , Imunossupressores/farmacologia , Prednisolona/farmacologia , Teste de Histocompatibilidade , Humanos , Teste de Cultura Mista de Linfócitos , Linfócitos T Citotóxicos/imunologiaRESUMO
Our standard procedure for phenotypic and functional analysis of immune cells present in the placenta is to isolate leukocytes from the decidua within five hours of the delivery. However, this results in logistical problems with deliveries at night, weekends or in other medical centers. Collecting placentas after complicated pregnancies is even more difficult owing to the low prevalence and the often unscheduled delivery. The aim was to investigate the possibility of preserving the human placenta before phenotypic and functional analysis of decidual lymphocytes. Placentas were obtained after uncomplicated pregnancy. The tissue was divided into two equal parts: decidual lymphocytes from one part were isolated within five hours according to our standard procedure, whereas the other part was preserved in either Celsior(®), a storage solution for solid organ preservation, or phosphate-buffered saline (PBS) for 24h at 4°C before isolation. Overall, the phenotype and functional capacity of decidual lymphocytes isolated within five hours was comparable to decidual lymphocytes isolated after 24-h preservation in Celsior(®) or PBS. Minor differences were found between decidual lymphocytes isolated within five hours and decidual lymphocytes isolated after 24-h preservation in Celsior(®). The results indicate that PBS is sufficient to preserve the placenta for 24h for phenotypical and functional studies. The ability to preserve the placenta will simplify the procedure for the isolation of decidual lymphocytes and makes it easier to analyze tissue from women who deliver during the night, at weekends or in other hospitals, and possibly even women with complicated pregnancies.
Assuntos
Linfócitos/imunologia , Preservação de Órgãos/métodos , Placenta/imunologia , Complicações na Gravidez/imunologia , Separação Celular , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Meios de Cultura/farmacologia , Dissacarídeos/farmacologia , Eletrólitos/farmacologia , Feminino , Citometria de Fluxo , Glutamatos/farmacologia , Glutationa/farmacologia , Histidina/farmacologia , Humanos , Imunidade Celular , Imunofenotipagem , Linfócitos/efeitos dos fármacos , Manitol/farmacologia , Estudos Multicêntricos como Assunto , Placenta/efeitos dos fármacos , GravidezRESUMO
A total of 64 loci for autosomal dominant non-syndromic hearing impairment have been described, and the causative genes have been identified for 24 of these. The present study reports on the clinical characteristics of an autosomal dominantly inherited hearing impairment that is linked to a region within the DFNA60 locus located on chromosome 2 in q22.1-24.1. A pedigree spanning four generations was established with 13 affected individuals. Linkage analysis demonstrated that the locus extended over a 2.96 Mb region flanked by markers D2S2335 and D2S2275. The audiograms mainly showed a distinctive U-shaped configuration. Deterioration of hearing started at a wide age range, from 12 to 40 years. Cross-sectional analysis showed rapid progression of hearing impairment from mild to severe, between the ages of 40 and 60 years, a phenomenon that is also observed in DFNA9 patients. The results of the individual longitudinal analyses were generally in line with those obtained by the cross-sectional analysis. Speech recognition scores related to the level of hearing impairment (PTA1,2,4 kHz) appeared to be fairly similar to those of presbyacusis patients. It is speculated that hearing impairment starting in mid-life, as shown by DFNA60 patients, could play a role in the development of presbyacusis. Furthermore, speech recognition did not deteriorate appreciably before the sixth decade of life. We conclude that DFNA60 should be considered in hearing impaired patients who undergo a rapid progression in middle age and are negative for DFNA9. Furthermore, cochlear implantation resulted in good rehabilitation in two DFNA60 patients.
Assuntos
Percepção Auditiva/genética , Cromossomos Humanos Par 2 , Genes Dominantes , Loci Gênicos , Perda Auditiva Neurossensorial/genética , Audição/genética , Adolescente , Adulto , Fatores Etários , Audiometria de Tons Puros , Audiometria da Fala , Criança , Implante Coclear , Progressão da Doença , Feminino , Predisposição Genética para Doença , Perda Auditiva Neurossensorial/diagnóstico , Perda Auditiva Neurossensorial/fisiopatologia , Perda Auditiva Neurossensorial/psicologia , Perda Auditiva Neurossensorial/reabilitação , Hereditariedade , Humanos , Masculino , Linhagem , Fenótipo , Reconhecimento Psicológico , Índice de Gravidade de Doença , Inteligibilidade da Fala , Percepção da Fala , Adulto JovemRESUMO
OBJECTIVE: To evaluate hearing impairment and cochlear function in non-ocular Stickler syndrome. STUDY DESIGN: Multifamily study. PATIENTS & METHODS: Ten patients from two different families with non-ocular Stickler syndrome (Stickler syndrome type 3) were included. Six members of the first family and four members of the second family participated in this study. Otorhinolaryngologic examinations were performed. Pure-tone and speech audiograms were obtained. Longitudinal analysis was performed. Psychophysical measurements, including loudness scaling, gap detection, difference limen for frequency and speech perception in noise were administered to assess cochlear function at a deeper level. RESULTS: Affected individuals in the first family were carriers of a heterozygous splice donor mutation in the COL11A2 gene. Affected individuals in the second family were carriers of a novel heterozygous missense mutation in COL11A2. Both families showed bilateral, non-progressive hearing impairment with childhood onset. The severity of the hearing impairment exhibited inter- and intrafamilial variability and was mostly mild to moderate. The results of the psychophysical measurements were similar to those previously published for DFNA8/12 (TECTA) and DFNA13 (COL11A2) patients and thus consistent with an intra-cochlear conductive hearing impairment. This is in line with the theory that mutations in COL11A2 affect tectorial membrane function. CONCLUSION: Hearing impairment in non-ocular Stickler syndrome is characterized by non-progressive hearing loss, present since childhood, and mostly mild to moderate in severity. Psychophysical measurements in non-ocular Stickler patients were suggestive of intra-cochlear conductive hearing impairment.