Killer cell immunoglobulin-like receptor expression delineates in situ Sézary syndrome lymphocytes

p140/KIR3DL2 has been identified in malignant cell lines isolated from the skin and blood of patients with transformed mycosis fungoides (MF) and Sézary's syndrome (SS). For the first time, the expression of a cell membrane structure appeared to be able to distinguish CD4+ tumour lymphocytes from reactive lymphocytes in these small cutaneous T-cell lymphomas (CTCLs). This study has examined the in vivo expression of this receptor in various CTCL subtypes, which constituted a heterogeneous group. Tumour cells diffusely expressed KIR in SS, in lymphomatoid papulosis (LyP) and in CD4+CD30+ as well as CD8+ large cell pleomorphic CTCL. In contrast, the infiltrating lymphocytes did not express KIR in MF at the patch/plaque stage or in CD4+CD30- large cell pleomorphic CTCL, except for scattered small cells. One quarter of the transformed MF tested exhibited KIR+ tumour cells, suggesting heterogeneity in this subtype. KIR expression was also examined in inflammatory lesions characterized by a dense infiltrate of T cells, such as lupus erythematosus and lichen planus. Only scattered CD8+ cells in lichen planus expressed a significant amount of KIR3DL2. Taken together, these results show for the first time that KIR molecules are expressed in distinct subtypes of malignant CTCL. It is also shown for the first time that SS and MF, which are frequent variants of CTCL with similar histological features, can be distinguished by their KIR3DL2 expression analysis. The identification of this KIR also differentiates between lupus erythematosus and lichen planus, which are both diseases with dense benign lymphocytic infiltrates.     

Killer cell immunoglobulin-like receptor (KIR) gene contents: Are they associated with cervical cancer?

Killer cell immunoglobulin-like receptors (KIRs) are required for natural killer cell function against virus-infected cells or tumor cells. KIR gene content polymorphisms in Indian women with cervical cancer (CaCx) remain unexplored. Hence, we analyzed the frequencies of KIR genes, KIR haplotypes, and Bx subsets to draw their association with CaCx. The polymerase chain reaction-sequence-specific primer method was used for KIR genotyping in three groups of women: healthy controls (n = 114), women with human papillomavirus (HPV) infection (n = 70), and women with CaCx (n = 120). The results showed that the frequency of KIR2DS5 was significantly higher in women with CaCx compared to women with HPV infection (p = 0.02) and healthy controls (p = 0.01). Whereas the frequency of KIR2DL5B was significantly higher in healthy controls than in women with HPV infection (p = 0.02). The total number of activating KIR genes was higher in women with CaCx than in healthy controls (p = 0.006), indicating their positive association with CaCx. Moreover, the C4T4 subset was higher in women with CaCx than in women with HPV infection, though not significant. In conclusion, our findings highlight KIR2DS5, the C4T4 subset, and activating KIR genes are susceptible factors or positively associated with CaCx. Besides KIR2DL5B, this study also reported for the first time significantly high frequency of KIR2DL1 in healthy controls, indicating its possible protective association against CaCx. Further, significantly high frequency of KIR2DL3 observed in HPV-infected women might be also a promising biomarker for viral infections. Thus, the study confirms the association of KIR genes with cervical cancer in women with HPV infection.     

HCV infection among cocaine users in the state of Pará, Brazilian Amazon

In this study, the prevalence, genotype frequency, and risk factors for HCV infection in 384 cocaine users were determined. One hundred twenty-four (32.3 %) cocaine users had anti-HCV antibodies and 120 (31.3 %) had HCV-RNA. Genotyping results indicated the predominance of genotypes 1 (73.3 %) and 3 (26.7 %). Multivariate analysis showed an association of HCV infection with tattoos, shared use of paraphernalia, daily cocaine use, and a long history of cocaine use. The epidemiological aspects of HCV infection among cocaine users presented here should serve as an incentive for the establishment of a program of hepatitis C prevention and control by the local public-health authorities in the Amazon.     

HLA-DRB1 and -DQB1 alleles in the Brazilian population of the northeastern region of the state of São Paulo

The HLA-DRB1 and -DQB1 alleles in 161 healthy unrelated individuals, including Caucasians, Blacks and Mulattos (mixed Caucasian and Black), from the Northeastern region of the state of S?o Paulo, Brazil were analysed. The 36 different DRB1 alleles detected included not only common Caucasian alleles, but also DRB1*0411, 0807 and 1402, typical of Amerindians, and DRB1*0302, 1503, and 0804, typical of African American Blacks.     

KIR and HLA ligands demonstrate genetic inheritance diversity in Japanese descendants from Paraná, Brazil

In order to investigate killer immunoglobulin-like receptors (KIR) and their ligands, human leukocyte antigen (HLA), diversity in the Brazilian population influenced by migrations, unrelated Brazilian Japanese descendants were selected and genotyped for the KIR genes and HLA class I allele groups. Genetic heterogeneity in Brazil Paraná Japanese was observed for KIR genes, whose frequency distributions demonstrated similarity with mixed Brazilian populations and with the Japanese population, suggesting gene flow. The data contributed to the identification of the genetic constitution of the Brazilian population influenced by immigrations and two new genotypes were defined.     

–308 G/A TNF-α gene polymorphism influences the course of basal cell carcinoma in a Polish population

Introduction

The etiopathogenesis of basal cell carcinoma (BCC) is multifactorial. The TNF-α gene seems to be an interesting gene candidate for BCC susceptibility because of the proinflammatory and immunosuppressive properties of its product. The aim of the study was to assess the frequency of –308 G/A and –238 G/A gene polymorphisms in the TNF-α gene and serum levels of cytokine in patients with BCC.

Material and methods

The study included 176 (94 women, 82 men) patients with BCC and 261 healthy volunteers. –308 G/A and –238 G/A TNF-α polymorphisms were analyzed using the amplification refractory mutation system-polymerase chain reaction method (ARMS-PCR). Serum concentrations of TNF-α were measured using ELISA.

Results

There was no statistically significant association between allele, genotype and haplotype frequencies in BCC patients in comparison with controls. Occurrence of the –308 TNF-α A allele or GA genotype in the group of patients with BCC increases risk of recurrence of tumor recurrence (OR = 4.8, 95% CI: 1.6–13.9, p = 0.004 and OR = 4.97, 95% CI: 1.7–14.5, p = 0.004). Moreover, –308 TNF-α GG genotype decreased risk of recurrence (OR = 0.2, 95% CI: 0.07–0.6, p = 0.004). The –238/–308 GA haplotype was connected with increased risk of recurrence (OR = 4.36, 95% CI: 1.49–12.7, p = 0.007). We also found significantly higher TNF-α levels among BCC patients in comparison with controls (p = 0.004).

Conclusions

The obtained results did not confirm the role of the –308 G/A and –238 G/A TNF-α gene polymorphisms in BCC development, but the presence of the A allele or GA genotype in –308 G/A TNF-α gene polymorphism may have an impact on the course of the disease.     

Immunity to flavivirus in Amerindian population of the Sierra de Perijá, Zulia state, Venezuela

Little information is available about Flavivirus infection in amerindian populations in western Venezuela. On this account the activity and seroprevalence of these viruses were determined and the hypothesis concerning the existence of a sylvatic cycle, conditioning the infection transmission of these viruses in indigenous populations, was studied. For this, blood samples from Yukpas (n=144) and Barí (n=110) communities were collected, 35 (Yukpas=25 and Barí=10) of which were processed for viral isolation followed by RT-PCR. The anti-Flavivirus IgG antibodies were determined by ELISA. The results did not show active Dengue cases and the seroprevalence of anti-Flavivirus IgG in the Yukpa population was significantly higher (p < 0.0001) than in the Barí population (43.1% vs. 6.4%). The present study has determined the presence of Flavivirus immunity in Yukpa and Barí populations. These results show a higher prevalence at the former than in the Barí population, which suggests circulation of Flavivirus, mainly in the Yukpa communities, being scarce and sporadic in Barí villages. However, in the indigenous populations studied, the causes or factors that determine the off set of Flavivirus infections in these zones could vary. The detected prevalence between both communities may be due to differences in the structure settlements and social habits. No evidences were found to support the presence of a sylvatic cycle in the Flavivirus transmission, specially of Dengue, in this population.     

Genotype analysis of the NF1 gene in the French Canadians from the Québec population.

We genotyped 19 NF1 families from the French Canadians of the Québec population with six intragenic polymorphic markers including 2 RFLPs (EcoRI and RsaI) and 4 microsatellites (IVS26-2.3, IVS27AC28.4, IVS27AC33.1, and IVS38GT53.0). Genotype analysis indicated families 7610 and 7473 bear deletions. In Family 7610 the deletion removed the entire NF1 gene except exons 1 to 4b. The breakpoint of the deletion is located between exons 4a and 4b. The deletion 7473 was derived from the maternal chromosome and exons 1 to 5 were deleted. The breakpoint of the deletion is located between exons 7 and 13. Their phenotypes are reported. The allele frequencies of microsatellites IVS27AC28.4 and IVS38GT53.0 are compared to previously reported data from Caucasians, including Spanish and Italians. The difference is statistically significant (P < 0.0036) for marker IVS27AC28.4 between the Québec French Canadian and the Italian population.     

Distribution of Δ32 alelle of the CCR5 gene in the population of Poland

The chemokine receptor CCR5 constitutes a major co-receptor for the R5 strains of HIV-1, and a mutant allele of the CCR5 gene, especially in the homozygous form Δ32/Δ32, confers resistance against infection by the virus. The frequency of the Δ32 allele was determined in blood donors from 16 provinces, covering the entire territory of Poland. Among 861 individuals 182 (21.1%) were carriers of the mutated allele; 7 of them (0.8 %) were homozygotes Δ32/Δ32, and 175 (20.3%) were heterozygotes +/Δ32, resulting in a 10.9% frequency of the Δ32 allele. The highest frequencies of the mutated allele were found in the eastern and western provinces, and the lowest frequencies of the Δ32 allele were detected in the provinces in the center of the country. This pattern of distribution may reflect the migration of the population from the eastern territories of Poland to the western part of the country after World War II. Received: March 17, 2000 / Accepted: May 29, 2000     

Isolation of the missing 5′-end of the encoding region of the bovine leukemia virus cell receptor gene

Summary The missing 5-end of the encoding region of the bovine leukemia virus (BLV) cell receptor gene (BLVRcp1/5) was isolated from a lambda gt11 cDNA library using the³²P-labeledEcoRI-SamI fragment corresponding to the 5-end of a 2.3 kbp cDNA fragment encoding the binding domain of the bovine leukemia virus cell receptor gene (BLVRcp1). The nucleotide and amino acid sequence analysis of the BLVRcp1/5 cDNA revealed that the 1058 bpEcoRI fragment at its 5-end contained a new 114 amino acid long sequence, and at its 3-end contained a completely identical 88 amino acid overlapping region with the 5-end of the BLVRcp1 cDNA. The combined sequences of both cDNAs represent the whole encoding region of the BLV cell receptor gene. The longest open reading frame of the BLV cell receptor gene encodes a protein containing 843 amino acids with a calculated molecular mass of 94.2 kDa which concurs with experimentally detected native BLV receptor protein. Search for homology has shown that about 250 bp of the BLV cell receptor gene is highly homologous to Venter's tag sequences of an unidentified gene from the human brain library.     

Hantaviruses in Central South America: phylogenetic analysis of the S segment from HPS cases in Paraná, Brazil

We sequenced the complete S segments of hantaviruses detected from 12 HPS patients living in southern of Brazil. Samples were obtained from patients diagnosed in different years, in distinct areas, and with a broad spectrum of clinical signs. Despite these differences, all the S proteins of hantavirus from Paraná were identical, except for one amino acid substitution. Phylogenetic analyses of the complete S segment nucleotide and amino acid sequences indicated that hantaviruses from Paraná form a distinct clade from those circulating in South and North America. Other hantaviruses from Brazil were not placed in the same clade. The Oligoryzomys nigripes-associated strains ITA37 and ITA38 from Paraguay were found to belong to the same clade as the hantaviruses from Paraná. Paraguay and Paraná state are located at the same latitude and some ecosystems are similar in both places. The geographic position and common rodent hosts could explain this phylogenetic relationship.     

Retinoic acid receptor β deficiency reduces splenic dendritic cell population in a conditional mouse line

Different studies have shown that retinoids and their receptors [retinoic acid receptors (RARs) and retinoid X receptors (RXRs)] have crucial effects on the differentiation and function of myeloid cells such as Dendritic cells (DCs) and the development of lymphoid tissue. However, the relationship between RARβ expression and DCs has not been previously studied in vivo. This work examined the effect of decreased RARβ expression on the number (and probably on differentiation) of splenic DCs and the structure of spleen using a conditional mouse that partially ablates floxed RARβ gene (RARβ(L-/L-) mice). Our results showed that RARβ is expressed mainly in cells of the splenic White Pulp (WP) zone of Wild type mice. As expected, low levels of RARβ expression were detected in the spleen of RARβ(L-/L-) conditional mice. These results were consistent with a decrease in the population of splenic CD11c(+)MHC-II(+) cells. Histopathological analyses of conditional mice spleen indicated defects in cell organization and structure. The expression of Toll-like receptor 2 was also down-regulated in the spleen of these mice. These results suggest that RARβ is involved in splenic cell organization as well as in the maintenance of splenic DCs population, indicating that RARβ expression is important in homeostasis of immune system components.     

Cervical cancer screening program of Paraná: cost-effective model in a developing country

The purpose of this study is to report the organization of a cost-effective screening program for cervical cancer in a developing country such as Brazil. The Cervical Cancer Screening Program of Paraná (CCSPP) was launched in October 1997 and was the result of a joint collaboration between the government of Paraná (Secretary of Health of the State of Paraná), scientific societies (pathologists, gynecologists, and nurses), and a non governmental organization called the Women's Popular Forum of Paraná. The main goal of the program was to enhance the Papanicolaou (Pap) smear screening to coverage up to 85% of female adult population with a 3-yr interval between examinations, as well as to reduce the incidence and mortality from cervical cancer in the state of Paraná, a Southern state of Brazil. The cytological findings in all Pap smears recorded in a central computer-based register during 5 yr of the program (October 1997-October 2002) are discussed. During that period, 2,244,158 Pap smears were performed in women included in the program from the 398 cities of the state of Paraná. The cytological smears were analyzed according to the Bethesda System. The previous year, before the program was launched, a Pap smear was taken from 43% of women of Paraná. At the end of 5 yr, coverage was increased to around 86%. The great majority of examinations had a negative result (98%). Only 2% of examinations had cytological abnormalities (n = 44,621). Low-grade lesions predominated in women aged 15-30 yr, and the high-grade lesions were more common in women aged 25-45 yr. Patients older than 40 yr had the greatest incidence of invasive cancer. Although the program is only 5 yr old, a decrease in the mortality from cervical cancer in women from Paraná is clearly apparent: in 1998, 297 women died of cervical cancer, as compared with 188 as of September 2002.     

γδ T cells represent a major spontaneously cytotoxic cell population in the chicken

Natural killer cells in the chicken are mainly confined to the intestine, while only small frequencies are detectable in spleen, lung and blood. Here, we compared the spontaneous cytotoxicity of lymphocytes isolated from blood, spleen and intestine using a flow cytometric based cytotoxicity assay. There was no spontaneous cytotoxicity detected in chicken blood preparations. In contrast, freshly prepared splenocytes exhibited a spontaneous cytotoxicity of up to 50% and intestinal epithelial lymphocytes of up to 85%. This cytotoxicity was observed against the RP9 but not against the chicken CU24 target cell line. The observed cytotoxicity was MHC unrestricted since B²B² derived effector cells killed RP9 target cells (B²B¹⁵) equally well compared to MHC mismatched 2D8 targets (B¹⁹B¹⁹). The cytotoxicity of splenocytes was enhanced by preincubation with IL-2 or strongly increased with IL-2 plus IL-12. By cell sorting, we identified the CD8⁺γδ T cell subset as the major effectors, whereas both CD8⁻γδ T cells and CD8⁺αβ T cells had only low cytolytic potential. Within intestinal lymphocyte CD45⁺cells displayed cytotoxicity as well as sorted γδ T cells and NK cell. In conclusion, the chicken γδ T cells represent a major cytotoxic lymphocyte subset that can lyse target cells in a MHC unrestricted manner.     

Extreme androgen resistance in a kindred with a novel insertion/deletion mutation in exon 5 of the androgen receptor gene

Androgen insensitivy syndrome (AIS) is the most frequent cause of male pseudohermaphroditism resulting from target-organ resistance to androgen action. Individuals bearing the complete form of the disease (CAIS) present a female phenotype and a lack of pubic and axillary hair. In the present study, four 46,XY patients born in two generations from a kindred with a history of AIS were examined for genetic abnormalities in the androgen receptor gene (AR). All eight exons encoding the AR protein were individually amplified from genomic DNA followed by a mutation screening with single-strand conformation polymorphism analysis. Sequencing of the mutant AR revealed a novel insertion/deletion mutation in exon 5. A deletion of 7 bp is replaced by an insertion of 11 nucleotides, which represents a duplication of the adjacent downstream sequence. The mutation g.2640_2646delAGGATGC/2652_2662insTTCGCCCCTGA, results in a frameshift that introduces a premature termination signal TGA, nine codons downstream. Such a rearrangement predicts a truncation of the AR, thereby deleting a large portion of the ligand-binding domain (amino acid position 768–919). Furthermore, although this mutation breaks the translational reading frame starting from codon 760, examination of the complementary DNA suggested that it does not disturb mRNA splicing. These changes have been found in all the patients and appear to account for the observed absence of detectable androgen binding to the AR in cultured fibroblasts and for the CAIS phenotype in the kindred. This disorder represents the first insertion/deletion mutation of the AR that probably arose by a slipped-strand mispairing mechanism.     

Relationship of IL-1 and TNF-α polymorphisms with Helicobacter pylori in gastric diseases in a Brazilian population

It is well known that the risk of development of gastric cancer (GC) in Helicobacter pylori-infected patients depends on several factors. Thus, the aim of this study was to investigate the effect of proinflammatory cytokine gene polymorphisms for IL-1β, IL-1RN and TNF-α on the development of GC in a Brazilian population. A total of 202 biopsies obtained from Brazilian patients with chronic gastritis and GC were included in the study. Infection with H. pylori cagA⁺ was determined by the polymerase chain reaction (PCR) as previously described. IL-1β, IL-1RN and TNF-α polymorphism genotyping was performed by restriction fragment length polymorphism PCR. Associations between gene polymorphisms, clinical diseases and virulence markers were evaluated using either the X² test or the Fisher exact test. Our results demonstrated that the IL-1β -511 C/C and IL-1β -511 C/T alleles were associated with chronic gastritis in H. pylori-positive patients (P = 0.04 and P = 0.05, respectively) and the IL-1β -511 C/C genotype was associated with GC (P = 0.03). The frequency of IL-1RN alleles from patients with chronic gastritis and GC indicated that there was no difference between the genotypes of the groups studied. Similar results were found for TNF-α -308 gene polymorphisms. Our results indicate that the IL-1β -511 C/C and C/T gene polymorphisms are associated with chronic gastritis and GC development in H. pylori-infected individuals.     

Is reduced CAG repeat length in androgen receptor gene associated with risk of prostate cancer in Indian population?

Shorter CAG repeats in androgen receptor (AR) gene have been found to be associated with an increased risk of prostate cancer (CaP). Ethnic variations in CAG repeat length may contribute to varying risks in different populations. To evaluate the prognostic significance of androgen receptor (AR) CAG repeats in Indian population for CaP, genomic DNA from 113 CaP, 57 benign prostate hyperplasia (BPH) patients and 133 normal healthy controls were examined by using a PCR-based GeneScan analysis. The mean number of CAG repeat in CaP was significantly lower as compared to the healthy controls (20.26 vs 22.98; p = 0.016). The odds ratio for CaP was 2.96 (p < 0.01), when individuals with short CAG repeat (< or =22) were compared with those having longer repeats (>22). A significant association was also observed between short CAG repeat and young age at diagnosis (OR 2.18; p = 0.04). The mean CAG repeat was not significantly different in BPH and healthy controls; however, BPH patients showed a tendency towards short CAG repeats. Thus, our results show that CAG repeat polymorphism in AR gene is significantly associated with CaP risk, suggesting that AR CAG polymorphism may act as a risk modifier to CaP in Indian population.