RESUMEN
Cystic fibrosis (CF) is a common recessive genetic disease caused by mutations in the gene encoding for the cystic fibrosis transmembrane conductance regulator (CFTR) protein. More than 1800 different mutations have been described to date. Here, we report 3 novel mutations in CFTR in 3 Italian CF patients. To detect and identify 36 frequent mutations in Caucasians, we used the INNO-LiPA CFTR19 and INNO-LiPA CFTR17+Tn Update kits (Innogenetics; Ghent, Belgium). Our first analysis did not reveal both of the responsible mutations; thus, direct sequencing of the CFTR gene coding region was performed. The 3 patients were compound heterozygous. In one allele, the F508del (c.1521_1523delCTT, p.PHE508del) mutation in exon 11 was observed in each case. For the second allele, in patient No.1, direct sequencing revealed an 11-base pair deletion (GAGGCGATACT) in exon 14 (c.2236_2246del; pGlu746Alafs*29). In patient No. 2, direct sequencing revealed a nonsense mutation at nucleotide 3892 (c.3892G>T) in exon 24. In patient No. 3, direct sequencing revealed a deletion of cytosine in exon 27 (c.4296delC; p.Asn1432Lysfs*16). These 3 novel mutations indicate the production of a truncated protein, which consequently results in a non-functional polypeptide.
Asunto(s)
Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Fibrosis Quística/genética , Mutación , Alelos , Secuencia de Bases , Fibrosis Quística/diagnóstico , Fibrosis Quística/patología , Análisis Mutacional de ADN , Exones , Femenino , Heterocigoto , Humanos , Lactante , Recién Nacido , Italia , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Índice de Severidad de la EnfermedadRESUMEN
The mechanisms by which hepatitis C virus (HCV) induces chronic infection in the vast majority of infected individuals are unknown. Sequences within the HCV E1 and E2 envelope genes were analyzed during the acute phase of hepatitis C in 12 patients with different clinical outcomes. Acute resolving hepatitis was associated with relative evolutionary stasis of the heterogeneous viral population (quasispecies), whereas progressing hepatitis correlated with genetic evolution of HCV. Consistent with the hypothesis of selective pressure by the host immune system, the sequence changes occurred almost exclusively within the hypervariable region 1 of the E2 gene and were temporally correlated with antibody seroconversion. These data indicate that the evolutionary dynamics of the HCV quasispecies during the acute phase of hepatitis C predict whether the infection will resolve or become chronic.
Asunto(s)
Evolución Molecular , Hepacivirus/genética , Hepatitis C Crónica/virología , Hepatitis C/virología , Proteínas del Envoltorio Viral/genética , Enfermedad Aguda , Adulto , Anciano , Anticuerpos Antivirales , Progresión de la Enfermedad , Femenino , Genes Virales , Variación Genética , Hepacivirus/inmunología , Hepacivirus/fisiología , Hepatitis C/inmunología , Anticuerpos contra la Hepatitis C/biosíntesis , Hepatitis C Crónica/inmunología , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , Estudios Prospectivos , Selección Genética , Factores de Tiempo , Proteínas del Envoltorio Viral/inmunología , Replicación ViralRESUMEN
The study aims were to assess the response of peripheral blood lymphocytes (PBL) of cancer patients to exogenous Interleukin 2 (IL 2) either by PHA-prestimulated or non PHA-prestimulated PBL, and to carry out preliminary experiments for a direct quantitative evaluation of endogenous IL 2 production by PBL cultures of cancer patients in order to define the actual role of IL 2 in the disease. Analysis of PBL subsets was also carried out with monoclonal antibodies in a selected group of patients. A total of 134 patients entered the study. Cancer sites were: larynx 32, breast 36, lung (NSC) 24, colorectal 17 and gynecologic 25. In the former 3 cancer sites staging showed localized or only locally advanced disease, and in the last 2 sites disseminated disease. Our results provided evidence that cancer patients exhibit a T-cell functional immunodepression, which progresses during tumor growth, so that the localized disease shows a low-grade defect, and advanced disease a high-grade defect. Our data also clearly suggested that the factor involved with a primary role in this functional immune impairment is the IL 2 deficiency. A perspective may be drawn on the therapeutic administration in vivo of IL 2 and IL 2-activated lymphokine-activated killer cells in controlled clinical trials of selected groups of cancer patients.
Asunto(s)
Interleucina-2/biosíntesis , Linfocitos/efectos de los fármacos , Neoplasias/inmunología , Células Cultivadas , Humanos , Células Asesinas Naturales/inmunología , Linfocitos/clasificación , Linfocitos/metabolismo , Fitohemaglutininas/farmacologíaRESUMEN
In order to determine whether the immunological abnormalities described in intravenous drug addicts (IDA), are due to HIV infection, other viral infections or to the abuse of narcotic drugs, we studied the T lymphocyte subsets and serological markers of infection with hepatitis B and delta virus, cytomegalovirus and Epstein Barr virus, in 49 IDA. The immunological and serological features of IDA were compared with the control group, made up of 20 healthy subjects. In intravenous drug abusers we found a significant increase in the number of total lymphocytes (P less than 0.01), T-lymphocytes (P less than 0.05), T-suppressor cells (P less than 0.05), and serum IgG levels (P less than 0.0001) as compared with the control group. The prevalence of serological markers of infection with hepatitis B virus, hepatitis delta virus, cytomegalovirus and Epstein Barr virus was significantly higher in IDA as compared with the controls. In conclusion our study demonstrates that T-lymphocyte subsets in IDA seronegative for HIV infection are characterized by an enhancement of peripheral lymphocyte cells with a normal OKT4/OKT8 ratio.
Asunto(s)
Seropositividad para VIH , Dependencia de Heroína/inmunología , Inmunoglobulinas/análisis , Linfocitos/inmunología , Virosis/inmunología , Adulto , Infecciones por Citomegalovirus/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Hepatitis B/inmunología , Hepatitis D/inmunología , Infecciones por Herpesviridae/inmunología , Herpesvirus Humano 4 , Humanos , Recuento de Leucocitos , Masculino , Linfocitos T/inmunología , Linfocitos T Reguladores/inmunologíaAsunto(s)
Interleucina-2/farmacología , Neoplasias Laríngeas/inmunología , Linfocitos/inmunología , Adulto , Anciano , Anticuerpos Monoclonales , Femenino , Humanos , Técnicas In Vitro , Interleucina-2/biosíntesis , Neoplasias Laríngeas/metabolismo , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Metástasis Linfática , Activación de Linfocitos/efectos de los fármacos , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Masculino , Persona de Mediana Edad , Mitógenos/farmacología , Fenotipo , Fitohemaglutininas/farmacología , Receptores Inmunológicos/efectos de los fármacos , Receptores Inmunológicos/inmunología , Receptores de Interleucina-2Asunto(s)
Adyuvantes Inmunológicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Mama/terapia , Extractos del Timo/uso terapéutico , Adulto , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/cirugía , Cisplatino , Femenino , Fluorouracilo , Humanos , Metotrexato , Persona de Mediana EdadRESUMEN
The aims of the investigation were 1) to determine if there are defects in interleukin-2 (IL-2) regulation on either phytohemagglutinin (PHA)-activated or non-PHA-activated peripheral blood mononuclear cells (PBMC) in cancer patients to ascertain the role of IL-2 in this disease; 2) to carry out preliminary experiments for a direct quantitative evaluation of endogenous IL-2 production by PBMC cultures; and 3) to evaluate the IL-2 receptor expression by PBMC of cancer patients. An assessment of lymphocyte subsets was also performed with monoclonal antibodies in a selected group of patients. A total of 170 patients entered the study. Cancer sites were larynx (48), breast (44), lung (30), colorectal(23), and gynecologic (25). Staging showed in the former three cancer sites predominantly localized or only locally advanced disease and in the latter two sites disseminated disease. PBMC cultures were performed with microtiter plate technique and 3H-thymidine uptake evaluation using polyclonal mitogens, IL-2, and a monoclonal antibody against IL-2 receptor. Our results provided evidence that the cancer patients exhibit a T-cell functional immunodepression, which, to some extent, progresses during tumor growth so that the localized disease shows a low-grade defect and advanced disease a high-grade defect. Our data also clearly suggest that IL-2 deficiency is the primary factor involved in this functional immune impairment. We found no significant defect in the IL-2 receptor expression by PBMC of cancer patients. Our data also seem to support the in vivo therapeutic administration of IL-2 and lymphokine-activated killer cells to cancer patients.
Asunto(s)
Interleucina-2/fisiología , Neoplasias/inmunología , Receptores de Interleucina-2/biosíntesis , Anticuerpos Monoclonales , Células Cultivadas , Femenino , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Activación de Linfocitos , Linfocitos/clasificación , Masculino , Persona de Mediana Edad , Fitohemaglutininas/farmacologíaRESUMEN
The aims of the investigation were: 1) to determine if there are defects in interleukin 2 (IL 2) regulation either on phytohemagluttinin (PHA)-activated or non PHA-activated peripheral blood mononuclear cells (PBMC) in cancer patients, in order to ascertain the role of IL 2 in this disease, and 2) to carry out preliminary experiments for a direct quantitative evaluation of endogenous IL 2 production by PBMC cultures of cancer patients. An assessment of lymphocytes subsets was also performed with monoclonal antibodies in a selected group of patients. A total of 159 patients entered the study. Cancer sites were: larynx, 49; breast, 42; lung (NSC), 25; colorectal, 18; and gynecologic, 25. In the former 3 cancer sites, staging showed localized or only locally advanced disease and in the later 2 sites it showed disseminated disease. Our results provided evidence that the cancer patients exhibit a T cell functional immunodepression, which progresses during tumor growth, so that the localized disease shows a low-grade defect and advanced disease shows a high-grade defect. Our data also clearly suggested that the factor involved with a primary role in this functional immune impairment is the IL 2 deficiency. In our study we have not found a substantial difference of activity between recombinant and nonrecombinant IL 2, although the comparison of the relative activities of the two types of IL 2 is not easy to make, since they are expressed in different ways; however the recombinant one appeared to be slightly more active, probably for the higher purity. Our data also seem to support the perspective of the in vivo therapeutic administration of IL 2 in cancer patients.
Asunto(s)
Interleucina-2/farmacología , Linfocitos/efectos de los fármacos , Neoplasias/inmunología , Fitohemaglutininas/farmacología , Neoplasias de la Mama/inmunología , Células Cultivadas , Neoplasias del Colon/inmunología , Femenino , Neoplasias de los Genitales Femeninos/inmunología , Humanos , Tolerancia Inmunológica , Técnicas In Vitro , Interleucina-2/biosíntesis , Neoplasias Laríngeas/inmunología , Neoplasias Pulmonares/inmunología , Linfocitos/clasificación , Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Linfocitos T/fisiologíaRESUMEN
Two familial cases of hairy cell leukaemia are reported: a daughter, 44-years-old, with a very unusual ultrastructural pattern found in hairy cells, the "tubuloreticular inclusions", and her mother, 71-years-old, who was affected six years later. Routine laboratory investigations, cytochemical and cytogenetic studies including HLA typing, as well as in vitro proliferative response of peripheral blood mononuclear cells (PBMC) to polyclonal mitogens and to exogenous interleukin 2, were performed. The immunological characterization by assessing the cell surface phenotypic markers with monoclonal antibodies and transmission electron microscopy (TEM) investigations were also carried out. In case 2 all tests were performed both on PBMC and on the bone marrow cells. To the best of our knowledge this is one of the first reports of such familial association. The possibility that genetic factors might play a role in the etiology of leukaemia in man is discussed: in our two cases, however, cytogenetic studies did not support this, while HLA typing revealed a non-significant association of HCL with DQw3 allele. Alternatively, an environmental factor has been considered, and a viral infection-perhaps by a retrovirus of the HTLV family has been suggested as tubuloreticular inclusions have been found in both hairy cell leukaemia, as reported, by us, and AIDS-LAS. However, a long time elapsed between the manifestation of HCL in the daughter and in the mother, and as the two patients had not been living together at that time, the possibility of a viral transmission seems minimal. The results of TEM and of immunological investigations are presented and discussed. Both, but particularly the latter, support the B cell nature of the hairy cell.
Asunto(s)
Leucemia de Células Pilosas/genética , Leucemia Linfocítica Crónica de Células B/genética , Adulto , Anciano , Médula Ósea/ultraestructura , Femenino , Estudios de Seguimiento , Humanos , Leucemia de Células Pilosas/patología , Leucemia Linfocítica Crónica de Células B/patología , Microscopía Electrónica , FenotipoRESUMEN
13 patients with Hodgkin's disease (HD) previously treated, 9 of whom were long-time (more than 2 yr) off-therapy, were studied for peripheral blood lymphocyte response to interleukin 2 and for lymphocyte subpopulations by means of in vitro cultures and monoclonal antibodies. The aim of the study was to ascertain the role played by interleukin 2 in the impaired cell-mediated immunity of HD patients. The results show a response of peripheral blood mononuclear cells of HD patients to either the T cell-specific polyclonal mitogens PHA and Con A or to the T cell-dependent, although B cell-specific, PWM, most significantly decreased compared to the normal response. As far as the interleukin 2 involvement in HD is concerned, our study suggests: an impaired endogenous interleukin 2 production by T lymphocytes, a most probable deficiency of the interleukin 2 receptor (Tac) expression and 3) a decrease of the number and/or of the function of NK cells no longer responsive in vitro to interleukin 2. The phenotypic analysis of peripheral blood mononuclear cells showed a slight decrease of total T cells (T3+), of the helper/inducer subset (T4+) and of the T4+/T8+ cells ratio. Our data seem to support the rationale for a therapeutical approach with interleukin 2 in controlled clinical trials also in HD patients, according to the experiments in progress in solid tumor patients.
Asunto(s)
Enfermedad de Hodgkin/sangre , Interleucina-2/farmacología , Linfocitos/efectos de los fármacos , Proteínas Recombinantes/farmacología , Adolescente , Adulto , Células Cultivadas , Femenino , Enfermedad de Hodgkin/tratamiento farmacológico , Humanos , Activación de Linfocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Mitógenos/farmacología , Linfocitos T/clasificación , Linfocitos T/efectos de los fármacos , Factores de TiempoRESUMEN
To study the spread of human immunodeficiency virus type 1 (HIV-1) in Sardinia, we conducted a multicentre prospective study of the prevalence of antibody to HIV-1 (anti-HIV-1) in various populations during 1985-1989. The highest anti-HIV-1 prevalence (61.4%) was found in intravenous drug users. Anti-HIV-1 was found in 32% of haemophiliacs, 4.2% of thalassemics and less than 1% in the other groups. We conclude that control of HIV infection in Sardinia will require a major expansion of prevention and treatment programs for drug addiction.
Asunto(s)
Infecciones por VIH/epidemiología , Seroprevalencia de VIH , VIH-1 , Adolescente , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Anticuerpos Anti-VIH/análisis , Infecciones por VIH/inmunología , VIH-1/inmunología , Hemofilia A/epidemiología , Humanos , Italia/epidemiología , Masculino , Estudios Prospectivos , Abuso de Sustancias por Vía Intravenosa/epidemiología , Talasemia/epidemiologíaRESUMEN
To investigate whether the human immunodeficiency virus (HIV) infection or the abuse of narcotic drugs or other viral infections may be responsible for immunologic abnormalities in parenteral drug abusers, sera from 168 consecutive individual patients were collected from 1985 to 1986. The sera were tested for antibody to HIV (anti-HIV), and the clinical, immunologic, and serologic characteristics of 83 seropositive and 53 seronegative parenteral drug abusers were compared. The presence of anti-HIV was significantly associated with a decreased number of T helper lymphocytes (P less than .001), a reduced T helper/suppressor ratio (P less than .001). Of the 83 seropositive patients, 63 (76%) had generalized lymphadenopathy and 16 (18%) had AIDS-related complex. No patient had AIDS. Parenteral drug abusers with AIDS-related complex had significant reductions in the number of T helper cells (P less than .01) and the T helper/suppressor ratio (P less than .01) compared with patients with lymphadenopathy syndrome (LAS), suggesting that parenteral drug abusers with HIV infection develop a progressive immunodeficiency. IgG antibody to cytomegalovirus was found in 75% of anti-HIV-positive and 45% of anti-HIV-negative parenteral drug abusers (P less than .01), but significant associations between anti-HIV and markers for other viruses were not found. Our data confirm that HIV infection is the major cause of low T helper cells and reversed T helper/suppressor ratio in parenteral drug abusers.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Seropositividad para VIH/inmunología , Trastornos Relacionados con Sustancias/inmunología , Linfocitos T/clasificación , Complejo Relacionado con el SIDA/inmunología , Adulto , Anticuerpos Antivirales/análisis , Citomegalovirus/inmunología , Humanos , Inyecciones , ItaliaRESUMEN
One of 4 siblings affected by hereditary spinocerebellar ataxia (HSCA) of Marie's type developed Hodgkin's disease (HD): the stage was IV B, the patient was submitted to conventional chemo- and radiotherapy and achieved complete remission. An accurate clinical, genetic and immunological study was carried out on all his family, including a complete HLA typing, a chromosome study, the immunophenotyping of peripheral blood mononuclear cells (PBMC), the PBMC response to polyclonal mitogens, to interleukin 2 (IL-2), to the association of PHA + IL-2 and the evaluation of the IL-2 receptor expression. No association was clearly demonstrable between an HLA haplotype and HSCA, while the patient with HSCA and HD was HLA-B18- and DQw3-positive (the last at homozygous level), two antigens known to be strongly associated with HD, mainly among the Sardinian ethnic group. The mode of inheritance of HD susceptibility is however completely different from that of Marie's HSCA. The chromosome study did not show any characteristic pattern of the karyotype, neither of the HSCA affected nor of the unaffected members. The immunological investigations did not elucidate any characteristic behavior of the family members, apart from the typical findings of HD seen on patients with HSCA and HD. Our study could not demonstrate any genetic and/or immunologic common background shared by the two diseases, HSCA and HD. Their coexistence in our patient, although the statistic probability is very low, seems to be a fortuitous coincidence more than the result of a common genetic and pathogenetic mechanism.
Asunto(s)
Enfermedad de Hodgkin/genética , Degeneraciones Espinocerebelosas/genética , Adulto , Anticuerpos Monoclonales , Diagnóstico Diferencial , Femenino , Antígenos HLA/análisis , Haplotipos , Enfermedad de Hodgkin/diagnóstico , Enfermedad de Hodgkin/inmunología , Humanos , Cariotipificación , Activación de Linfocitos , Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Fenotipo , Receptores de Interleucina-2/inmunología , Degeneraciones Espinocerebelosas/diagnóstico , Degeneraciones Espinocerebelosas/inmunologíaRESUMEN
BACKGROUND AND METHODS: Chronic hepatitis D is a severe and rapidly progressive liver disease for which no therapy has been proved effective. To evaluate the efficacy of treatment with interferon, we studied 42 patients with chronic hepatitis D who were randomly assigned to receive either 9 million or 3 million units of recombinant interferon alfa-2a (three times a week for 48 weeks) or no treatment. RESULTS: By the end of the treatment period, serum alanine aminotransferase values had become normal in 10 of 14 patients receiving 9 million units (71 percent), as compared with 4 of 14 treated with 3 million units (29 percent, P = 0.029) and 1 of 13 untreated controls (8 percent, P = 0.001). Seven patients treated with the higher dose of interferon (50 percent) had a complete response (normal levels of alanine aminotransferase and no detectable serum hepatitis delta virus [HDV] RNA), as compared with three of those who received the lower dose (21 percent, P = 0.118), and none of the controls (P = 0.004). Treatment with 9 million units of interferon was associated with a marked improvement in the histologic findings (reduced periportal necrosis and portal and lobular inflammation), whereas in the untreated controls there was considerable histologic deterioration. In 5 of the 10 patients treated with 9 million units of interferon whose alanine aminotransferase values became normal, the biochemical responses persisted for up to 4 years (mean, 39 months), but the effects of treatment on viral replication were not sustained. In contrast, none of those who received 3 million units and none of the untreated controls had a sustained biochemical or virologic response. CONCLUSIONS: In about half the patients with chronic hepatitis D treated with high doses of interferon alfa-2a (9 million units three times a week for 48 weeks), the serum alanine aminotransferase level becomes normal, HDV RNA becomes undetectable in serum, and there is histologic improvement. However, a relapse is common after treatment has been stopped.
Asunto(s)
Hepatitis D/terapia , Interferón-alfa/uso terapéutico , Adolescente , Adulto , Alanina Transaminasa/sangre , Enfermedad Crónica , Femenino , Estudios de Seguimiento , Hepatitis D/enzimología , Hepatitis D/patología , Humanos , Interferón alfa-2 , Interferón-alfa/administración & dosificación , Interferón-alfa/efectos adversos , Hígado/patología , Masculino , Persona de Mediana Edad , Necrosis , Proteínas RecombinantesRESUMEN
Hepatitis C virus (HCV) was transmitted from a patient with fulminant hepatitis C to a chimpanzee. The patient had developed two episodes of fulminant hepatitis C, each occurring after a separate liver transplantation. Serial serum and liver samples from the patient and the chimpanzee were analyzed for HCV replication, genotype, quasispecies heterogeneity, and antibodies. In the patient, the levels of HCV replication in serum and liver correlated with the degree of hepatocellular necrosis and the clinical expression of fulminant hepatitis. The same HCV strain, genotype 1a, was recovered from both episodes of fulminant hepatitis. An unusually severe acute hepatitis was also observed in the chimpanzee. The viruses recovered from the patient and the chimpanzee were almost identical and displayed relatively little quasispecies heterogeneity. Thus, the same HCV strain induced two episodes of fulminant hepatitis in a single patient and severe hepatitis in a chimpanzee, suggesting that the pathogenicity or virulence of a specific HCV strain may be important in the pathogenesis of fulminant hepatitis C.