MHC2TA rs4774C and HHV-6A active replication in multiple sclerosis patients.

BACKGROUND AND PURPOSE: In a previous report, a strong gene-environment interaction between human herpesvirus 6A (HHV6A) active replication and MHC2TA rs4774C was demonstrated. The objectives of this study were: (i) to reappraise the association that was found in the previous study; (ii) to evaluate if MS patients with minor allele C and HHV-6A active infection had different clinical behavior; and (iii) to analyze the possible association of MHC2TA rs4774C with Epstein-Barr virus (EBV). METHODS: A total of 149 MS patients were analyzed both at the MHC2TA locus and by HHV-6A status in serum. We studied a G/C polymorphism (rs4774) by a TaqMan Assay-on-Demand. HHV-6A genomes in serum were evaluated by quantitative PCR. A control group of 562 healthy Spanish individuals was included for comparative purposes in the genetic analyses. A battery of clinical data was collected for all the MS patients included in the study. RESULTS: (i) MHC2TA/HHV-6A interaction: we found the same strong association of the rs4774C allele with HHV-6A active replication than in the previous study (P = 0.0001). (ii) CLINICAL DATA: the two main statistical significant differences for MS patients with HHV-6A active infection and minor allele C were: (a) a significant number of them were not free of progression (EDSS = 0) 2 years after the diagnosis (P = 0.01); (b) only a third of them responded to interferon beta treatment (P = 0.05). CONCLUSIONS: This study has verified previous results about the strong gene-environment interaction between HHV6A active replication and MHC2TA rs4774C. Furthermore, a different clinical behavior for MS patients with HHV-6A active infection and minor allele C was found.

Papel de los polimorfismos del promotor del gen de la interleucina-10 en la génesis de los aneurismas de la aorta abdominal / Role of polymorphisms in the interleukin-10 gene promoter in the development of an abdominal aortic aneurysm

Introducción. La interleucina-10 (IL-10) es una citocina antiinflamatoria que se ha implicado en la patogenia de los aneurismas de la aorta abdominal (AAA). Se han descrito variaciones en el promotor de su gen que pueden condicionar su producción, y alterar el equilibrio entre citocinas pro y antiinflamatorias. Objetivo. Estudiar el genotipo de los microsatélites y polimorfismos del promotor de la IL-10 para valorar las posibles predisposiciones genéticas en la población española portadora de un AAA. Sujetos y métodos. Se realizó el tipado de los microsatélites asociados (IL-10R e IL-10G) y de las tres posiciones polimórficas (-1082, -819, -592) del promotor de la IL-10 en un grupo de 72 pacientes con un AAA, y se comparó con una muestra control de 349 sujetos sanos. Resultados. Microsatélites asociados: no se encontraron diferencias significativas al analizar el patrón fenotípico de los microsatélites IL-10G e IL-10R entre los pacientes con un AAA y el grupo control. Polimorfismos puntuales: no se encontraron diferencias significativas al analizar los diferentes haplotipos del promotor del gen de la IL-10 entre los pacientes con un AAA y el grupo control. Conclusiones. A la vista de nuestros resultados, en la población española no existe una relación entre ninguno de los diferentes alelos de los microsatélites IL-10R e IL-10G asociados al promotor de la IL-10 y la presencia de un AAA en dicha población. Tampoco existe una relación entre los diferentes haplotipos del promotor de la IL-10 y esta enfermedad en nuestro medio

INTRODUCTION. The anti-inflammatory cytokine interleukin-10 (IL-10) has been implicated in the pathogenesis of the abdominal aortic aneurysm (AAA). Changes in the promoter region of the IL-10 gene have been found to condition its production, altering the balance between pro- and anti-inflammatory cytokines. AIM. Associated microsatellites and polymorphisms in the IL-10 promoter were typed in an attempt to identify a possible genetic predisposition for AAA in the Spanish population. SUBJECTS AND METHODS. Associated microsatellites (IL-10R and IL-10G) and the three polymorphic loci (-1082, -819, -592) of the IL-10 promoter were determined in 72 patients with AAA and compared with those established in 349 healthy control subjects. RESULTS. No significant differences were observed between the AAA and control group in the phenotypic patterns shown by the microsatellites IL-10G and IL-10R nor in the different haplotypes of the IL-10 gene promoter. CONCLUSIONS. Our findings suggest no relationship between the different alleles of the IL-10R and IL-10G microsatellites and AAA in the Spanish population. We were also unable to detect a clear association between the different IL-10 promoter haplotypes and this disease

A functional variant in the CD209 promoter is associated with DQ2-negative celiac disease in the Spanish population.

AIM: To address the role of CD209 in celiac disease (CD) patients. Non-human leukocyte antigen (HLA) genetic factors in CD predisposition are poorly understood, and environmental factors like infectious pathogens may play a role. CD209 is a dendritic and macrophage surface molecule involved in pathogen recognition and immune activation. Recently, a functional variant in the promoter of the CD209 gene (-336A/G) has been shown to affect the transcriptional CD209 activity in vitro and it has been associated with a higher susceptibility to/or severity of infection. METHODS: The study population was composed of two case-control cohorts of 103 and 386 CD patients and 312 y 419 healthy controls as well as a panel of 257 celiac families. Genotyping for the -336A/G CD209 promoter polymorphism was performed using a TaqMan 5' allelic discrimination assay. HLA-DQ was determined by hybridization with allele specific probes. RESULTS: Initially, the case-control and familial studies did not find any association of the -336 A/G CD209 genetic variant with CD susceptibility. However, the stratification by HLA-DQ2 did reveal a significant association of CD209 promoter polymorphism in the HLA-DQ2 (-) group (carrier A vs GG in DQ2 (-) vs DQ2 (+) patients (P = 0.026, OR = 3.71). CONCLUSION: The -336G CD209 allele seems to be involved in CD susceptibility in HLA-DQ2 (-) patients. Our results might suggest a possible role of pathogens in the onset of a minor group of CD patients.

A functional variant of IFNgamma gene is associated with coeliac disease.

In coeliac disease (CD) a profile of proinflammatory cytokines are secreted interferon gamma (IFNgamma) being one of the most important. A dinucleotide polymorphism consisting of a variable number of CA repeats related with IFNgamma production levels, has been reported on the first intron of the IFNgamma gene. The aim of this study was to analyse the influence of the functional IFNgamma CA repeats in CD predisposition through familial and case-control studies. The familial analysis showed that the 124 bp allele was significantly more transmitted to the affected offspring (P=0.02), while the 126 bp allele showed a statistically significant nontransmission pattern (P=0.01). Nevertheless, in the case-control analysis, we could not find a direct association of CA repeats with CD. This fact might be due to parent-of-origin effect in the IFNgamma CA polymorphism. Our data suggest a possible role of IFNgamma CA polymorphism in CD susceptibility.

[Basal immunological parameters in a group of retirees]. / Parámetros inmunológicos basales en un grupo de viejos-jóvenes.

OBJECTIVES: a) establish the major immunological parameters for clinical use in a group of retirees; b) correlate its levels in relation to gender; c) assess the influence of some specific factors (substance abuse, diseases) on the analyzed parameters. MATERIAL AND METHODS: study period: 1990-1999; sample: 249; 102 men (M); 147 women (W). Median age: 67.03 (4.2) years. Analyzed immunological variables: total leukocytes, lymphocytes B, immunoglobulins (IgG, IgA and IgM), rheumatoid factor, lymphocytes subpopulations (CD4, CD8, ratio), natural killer, complement (C3 and C4) and delayed hypersensitivity tests. TECHNIQUES: flow cytometry (EPICS-Profile II) and Multitest IMC. STATISTICAL ANALYSIS: SPSS version 10.0.Results. Gender influence: leukocytes: M: 6,699.4 (1,615.0); W: 6,105.9 (1,470.5); p < 0,003; lymphocytes B (%): M: 9.4 (5.0); W: 11.3 (4.1); p < 0,003; IgG: M: 1,155.7 (320.0); W: 1,116.1 (257.8); p = 0,28, IgM: M: 112.7 (69.3); W: 136.8 (85.6); p < 0,01; IgA: M: 276.1 (114); W: 254.0 (122); p = 0,15; rheumatoid factor: M: 18.5 (6.6); W: 20.9 (18.8); p = 0,020; CD4 (%): M: 42.2 (9.7); W: 47.3 (9,1); p < 0.001; CD8 (%): M: 30.3 (10,8); W: 25.0 (10.2); p < 0,001; scores: M: 13.2 (7.4); 11m: 10.0 (7.2); p < 0,005. Influence of the substance abuse: smokers; lymphocytes B (%): 8.8 (3.4); No: 10.9 (4.7); p < 0.008; CD8 (%): smokers: 31.8 (13.2); No: 26.2 (9.9); p < 0.003; CD4/CD8 ratio: smokers: 1.6 (0.9); No: 2.0 (1.3); p < 0,05; scores: smokers: 14.3 (6.8); No: 10.8 (7.5); p < 0.02; alcoholism: lymphocytes B (%): 8.7 (2.5). No: 10.8 (4.7); p < 0.001; alcoholism: store: 16.9 (6.7); No: 10.7 (7.3); p < 0,001. Influence of the diseases: diabetes: CD4 (%): 49.4 (12.0); diabetics versus healthy: p = 0,05; CD4/CD8 ratio: 2.6 (2.5); diabetics versus healthy: p = 0,04; EPOC: CD8 (%): 32.9 (16,3); COPD versus healthy: p = 0,07; neoplasias NK (%): 17.1 (21.4); neoplasias versus healthy: p < 0,01. CONCLUSIONS. a) there are differences according to the gender in the parameters of normalcy of some variables; b) smoking and alcoholism alter the immunological test analyzed, and c) some chronic diseases influence the subpopulations of lymphocytes and the cutaneous test of delayed hypersensitivity.

Parámetros inmunológicos basales en un grupode viejos-jóvenes / Basal inmunological parameters in a group of retirees

Objetivos. a) valorar los principales parámetros inmunológicos de uso clínico en un grupo de jubilados; b) correlacionar sus niveles en función del género, y c) valorar la influencia sobre los parámetros analizados de determinados factores (hábitos tóxicos, enfermedades).Material y métodos. Período de estudio: 1990-1999; muestra: 249; 102 hombres (H); 147 mujeres (M). Edad media: 67,03 ñ 4,2 años. Variables inmunológicas analizadas: leucocitos totales, linfocitos B, inmunoglobulinas (IgG, A y M), factor reumatoide, subpoblaciones de linfocitos (CD4, CD8, ratio), natural killer (NK), complemento (C3 y C4) y pruebas de hipersensibilidad retardada. Técnicas: citometría de flujo (EPICS-Profile II) y Multitest IMC. Análisis estadístico: SPSS versión 10.0.Resultados. Influencia del género: leucocitos: H: 6.699,4 (1.615,0); M: 6.105,9 (1.470,5); p< 0,003; linfocitos B ( por ciento); H: 9,4 (5,0); M: 11,3 (4,1); p < 0,003; IgG; H: 1.155,7; (320,0) M: 1.116,1 (257,8); p = 0,28; IgM; H: 112,7 (69,3); M: 136,8 (85,6); p < 0,01; IgA; H: 276,1 (114); M: 254,0 (122); p = 0,15; factor reumatoide: H: 18,5 (6,6); M: 20,9 (18,8); p = 0,20; CD4 ( por ciento); H: 42,2 (9,7); M: 47,3 (9,1); p < 0,001; CD8 ( por ciento); H: 30,3 (10,8); M: 25,0 (10,2); p < 0,001; score; H: 13,2 (7,4); M: 10,0 (7,2); p < 0,005. Influencia de los hábitos tóxicos: fumadores: linfocitos B ( por ciento): 8,8 (3,4); No: 10,9 (4,7); p < 0,008; CD8 ( por ciento); fumadores: 31,8 (13,2); No: 26,2 (9,9); p < 0,003; cociente CD4/CD8: fumadores: 1,6 (0,9); No: 2,0 (1,3); p < 0,05; score: fumadores: 14,3 (6,8); No: 10,8 (7,5); p < 0,02; hábito enólico, linfocitos B ( por ciento): 8,7 (2,5); No: 10,8 (4,7); p < 0,001; hábito enólico, score: 16,9 (6,7); No: 10,7 (7,3); p < 0,001. Influencia de las enfermedades: diabetes; CD4 ( por ciento): 49,4 (12,0); diabéticos frente a sanos: p = 0,05; cociente CD4/CD8: 2,6 (2,5); diabéticos frente a sanos: p = 0,04; enfermedad pulmonar obstructiva crónica (EPOC): CD8 ( por ciento), 32,9 (16,3); EPOC frente a sanos: p = 0,07; neoplasias: NK ( por ciento): 17,1 (21,4); neoplasias frente a sanos: p < 0,01. Conclusiones. a) existen diferencias en función del género en los parámetros de normalidad de algunas variables; b) el tabaquismo y el hábito enólico alteran las pruebas inmunológicas analizadas, y c) determinadas enfermedades crónicas influyen en las subpoblaciones de linfocitos y en las pruebas cutáneas de hipersensibilidad retardada (AU)

Papel del hla en la patogenia de los aneurismas de aorta abdominal / Role of hla complex in the pathogenesis of abdominal aortic aneurysms

Introducción. En los últimos años, se han descrito alteraciones en el patrón de las metaloproteinasas en los aneurismas de aorta abdominal (AAA), secundarias a procesos inflamatorios que podrían estar implicados en la patogenia de los mismos; existen muy escasos estudios que valoren el papel de la autoinmunidad en la etiopatogenia de esta enfermedad. Objetivo. Estudiar el patrón HLA de clase II y III para valorar la participación de un proceso autoinmune en los AAA. Pacientes y métodos. Se realizó el tipaje de los HLA-II y las zonas del HLA-III correspondientes al factor de necrosis tumoral (TNF) y su promotor en un grupo de 72 pacientes con AAA, y se comparó con una muestra control de 380 personas sanas. Resultados. HLA-II: se encontró mayor incidencia de HLADR-4*401 en AAA respecto a controles (12,5 por ciento frente a 5,2 por ciento; p= 0,02; OR= 2,59). No hubo diferencias entre los demás alelos. TNF: la frecuencia de aparición del TNF a4b5 en el grupo de AAA mostró una tendencia a la significación (p= 0,055) sin encontrar otras diferencias respecto a los controles. Tampoco se encontraron diferencias en cuanto a los promotores del TNF. Haplotipos: el haplotipo HLA-DR3-TNF a2b3 fue significativamente más prevalente entre los AAA respecto a los controles (16,6 por ciento frente a 8,4 por ciento; p= 0 ,03; OR= 2,17). Conclusiones. La relación encontrada entre el HLA-DR4*401 y el haplotipo ancestral HLA-DR3-TNF a2b3 que se encuentra asociado a otras enfermedades autoinmunes, sugiere que fenómenos de autoinmunidad pueden desempeñar un papel importante en la patogenia de los aneurismas de aorta abdominal. (AU)

[Genetics of autoimmune diseases]. / Genética de las enfermedades autoinmunes.

The publication of the human genome sequence this year was an important milestone in Biology. Autoimmune diseases arise through a complex interaction of genetics and environmental factors. They are influenced by more than one gene and do not exhibit a simple mode of inheritance. Some of the involved genes are thought to play a role in several autoimmune diseases whereas others seem to be specific for one of them. We have studied mainly genes located on the Major Histocompatibility Complex situated on the short arm of the 6th chromosome. This region is the most important genetic determinant for autoimmune diseases. It carries many genes playing a role in immune response and several have been observed to be associated with autoimmune diseases susceptibility. Although what has been published is just a draft of the human genome sequence, and a lot of work is needed to make sense of all these data and move into the study of gene function, the important technology breakthroughs of the last few years in genomics, proteomics and bioinformatics will speedup the study and results may come much earlier than could be imagined some years ago.

Influenza virus immunization effectivity in kidney transplant patients subjected to two different triple-drug therapy immunosuppression protocols: mycophenolate versus azathioprine.

BACKGROUND: Due to possible complications and treatment limitations, the prevention of influenza in renal transplant (RT) patients is highly indicated. METHODS: Forty-nine patients with a 1-year functioning RT subjected to two different immunosuppressive regimens and 37 healthy relatives (HR) were administered the anti-influenza vaccine as recommended for 1996 to 1997. Anti-influenza antibody, creatinine, and immunological markers were estimated at 1 and 3 months after vaccination. RESULTS: Three months after vaccination, 46.2% of the RT patients and 69% of the HR (P=0.06) showed protective antibody titers to influenza A (relative risk [RR]; 0.67; 95% confidence interval: 0.44-1.02). A total of 20.5% of the RT patients and 44.8% of the HR showed antibodies to influenza B (P=0.03). Despite these differences, the incidence of illness was similar. The immunosuppressive regimen had no effect on the antibody response. CONCLUSIONS: Although the RT patients showed a reduced antibody response, no negative effects on graft outcome were observed.

Importancia del factor de necrosis tumoral en la enfermedad celíaca / Importance of tumor necrosis factor in coeliac disease

Las asociaciones de los microsatélites de TNF a y b con la susceptibilidad a la enfermedad celíaca (EC), han sido estudiadas en 137 pacientes y 324 controles. Nuestros datos indican una asociacíón entre EC y el haplotipo a2b3 del factor de necrosis tumoral (AU)

LOH at the APC/MCC gene (5Q21) in gastric cancer and preneoplastic lesions. Prognostic implications.

The APC/MCC gene (Familial Adenomatous Polyposis) at 5q21 plays a role in colon cancer carcinogenesis. LOH at this locus has also been described in gastric cancer and preneoplastic lesions. The APC locus has been recently related to a cell surface adhesion molecule and its alteration may favour metastatic dissemination. LOH at 5q21 has been associated with poor prognosis in other tumors such as lung cancer. Thirty-six gastric cancers were evaluated for LOH at 5q21 with 2 polymorphic markers from microdissected paraffin-embedded material. All tumors were classified by stage, histologic type, degree of differentiation and survival rates. In 4 cases, intestinal metaplasia cells in the adjacent mucosae were also microdissected. Six cases of moderate-severe gastric dysplasia were also added to the study. LOH was determined in 84% of the informative cases of GC, affecting both early and advanced stages of disease. Genomic instability was assessed in 5 cases, 3 of them associated with LOH. The only case of gastric cancer that did not show LOH or instability at 5q21 was a stage II, poorly differentiated intestinal carcinoma without evidence of recurrence after a 36 month follow-up period (the mean survival rate in our series was 28.3% at 36 months). We also found LOH in 2/6 dysplastic lesions and 1/4 intestinal metaplasias. Our data show that LOH at 5q21 is frequent in gastric cancer and is also present in intestinal metaplasia and dysplastic lesions. LOH at this locus is not a prognostic factor in GC in our study, due to the high incidence of LOH that we found.

Increased serum levels of IgA antibodies to hsp70 protein in patients with diabetes mellitus: their relationship with vascular complications.

We measured class-specific antibodies to the mycobacterial hsp70 protein in 67 patients with diabetes mellitus (27 type 1 and 40 type 2) with or without vascular complications. Using ELISA, the levels of IgG and IgM antibodies in the sera of diabetic patients did not significantly differ either from those of healthy control subjects or between both types of diabetes, regardless of gender, disease duration, HbA1 level, or type of vascular complication. In patients with type 2 diabetes, the mean serum IgA levels were significantly higher than those in their matched controls [274(71) mg/dl vs 208(88) mg/dl; P < 0.01]. In this group of patients, the IgA antibody titer was significantly correlated to the serum IgA level (r = 0.334; P < 0.01). Serological autoimmunity (IgG or IgM type) to hsp70 protein is common in both the normal and the diabetic population. The increased IgA levels and anti-hsp70 IgA titers in the sera of diabetics suggest a possible role of IgA in the pathogenesis of the vascular complications of diabetes mellitus.

[Lymphocyte subpopulations and surgery. The role of postoperative parenteral nutrition]. / Subpoblaciones linfocitarias y cirugía. Papel de la nutrición parenteral postoperatoria.

The effects of LCT-based lipid emulsions used in TPN on immune system remains controversial. In this prospective study we research the effects of three types of TPN on T-lymphocyte subsets and NK cells. 21 patients diagnosed because of upper gastrointestinal carcinoma (UGIC), and amenable of curative surgery were included in the study. TPN support was maintained 10 postoperative days at least. All patients received 35 non-proteic Kcal/KG BW/day. Group I (without lipid): received 100% of caloric intake (CI) by glucose. Group II (LCT): received 55% of CI by glucose and 45% by LCT at 20% emulsion. Group III (MCT/LCT): received 55% of CI by glucose and 45% by MCT/LCT at 20% mixture. T-lymphocyte subsets were determined by flow cytometry preoperatively and in first and tenth postoperative days. Our results suggest that patients diagnosed of UGIC present alterations of cellular immunity. These alterations are increased by the age and by surgical act. The changes found are independent of the type of TPN. LCT-based emulsions have similar effects on T-lymphocyte subsets that MCT/LCT-based emulsions.

[Lymphocyte subpopulations in the evolution of brucellosis]. / Subpoblaciones linfocitarias en la evolución de la brucelosis.

We have sequentially studied for one year CD3+, CD4+, CD8+, CD11b+, and CD20+ lymphocyte subpopulations in 52 brucellosis patients whose diagnosis was confirmed by hemoculture. We observe at diagnosis a decreased percentage of CD4+ lymphocytes with an increase in CD8+ and inverted CD4+/CD8+ ratio, which was greater in patients with an evolution longer than 4 weeks (p less than 0.05) returning to normal very slowly. The percentage of monocytes was also increase at diagnosis (p less than 0.01) and at their second month CD3+ an CD20+ subpopulations did not suffer any changes during the follow up period. Patients were divided into two groups according to whether one or two anti-Brucella IgG peaks were observed in ELISA throughout the evolution. Patients in whom IgG titer decreased uninterruptedly, the inversion of the ratio during the first two months was greater, while those patients who suffered a relapse showed a simultaneous new decrease in CD4/CD8 ratio (p less than 0.05).