ITPKC and SLC11A1 Gene Polymorphisms and Gene-Gene Interactions in Korean Patients with Kawasaki Disease

PURPOSE: Kawasaki disease (KD) is an acute systemic vasculitis. Both the etiology of KD and the erythema of Bacille Calmette-Guérin (BCG) injection sites observed in the disease are poorly understood. We investigated the association between KD and single nucleotide polymorphisms (SNPs) in two candidate genes: inositol 1,4,5-triphosphate 3-kinase (ITPKC), a well-studied KD-associated gene, and solute carrier 11a1 (SLC11A1), which is associated with the hypersensitive reaction to the BCG strain in Koreans. MATERIALS AND METHODS: Associations between KD and SNPs in two genes were evaluated. Potential associations between BCG injection site erythema and SNPs in two genes were also evaluated. Gene-gene interactions between ITPKC and SLC11A1 in KD and BCG injection site erythema were also analyzed. RESULTS: Three tagging SNPs in ITPKC and five tagging SNPs in SLC11A1 were genotyped in 299 KD patients and 210 control children. SNP rs28493229 in ITPKC was associated with KD and coronary artery complications. SNP rs77624405 in SLC11A1 was associated with KD. Comparisons of KD patients with and without BCG injection site erythema revealed that SNP rs17235409 in SLC11A1 was associated with erythema; no erythema-associated SNPs in ITPKC were identified. Interactions between ITPKC rs28493229_GG and SLC11A1 rs17235409_GA and between ITPKC rs10420685_GG and SLC11A1 rs17235409_AA were strongly associated with BCG injection site erythema. CONCLUSION: This study identified several important polymorphisms in the ITPKC and SLC11A1 genes in Koreans. The genetic variants identified in this study affected KD and erythema of BCG injection sites independently and through gene-gene interactions. Also, the effects of the polymorphisms were age-dependent.

Role of the gene Slc11a1 and selected genotypes for minimum and maximum inflammatory reactivity in the process of alveolar bone healing in mice / Papel do gene Slc11a1 e de genótipos selecionados para mínima e máxima reatividade inflamatória no processo de reparo ósseo alveolar em camundongos

The process of alveolar bone healing can be influenced by several local and systemic factors, which include the immune system and healing related genes. However, the exact role of host inflammatory responsiveness and genetic background in bone healing process remains unclear. In this context, we evaluated the influence inflammation in alveolar bone healing taking advantage of mice strains genetically selected for maximum (AIRmax) or minimum (AIRmin) acute inflammatory response, as well AIR strains homozygous for RR/SS Slc11a1 genotypes. Experimental groups (N=5/time/group) comprised 8-week-old male or female AIRmax and AIRmin; and substrains AIRminRR, AIRminSS and AIRmaxRR and AIRmaxSS; submitted to extraction of upper right incisor and evaluated at 0, 3, 7, 14 and 21 days after upper incision extraction by micro-computed tomography (CT), histomorphometry, birefringence, immunohistochemistry and molecular (PCRArray) analysis. Initially, our results demonstrated that AIRmin mice presented an early increase (p<0.05) in bone volume, hyperdense regions, density of bone matrix and osteoblasts, increased (p<0.05) expressed of BMP4, BMP7 and RUNX2 when compared to AIRmax strain. AIRmin mice also presented lower counts of GR1+ and CD80+ cells, and higher counts of F4/80+ and CD206+ cells, in parallel with higher mRNA expression of CX3CL1, CCL5, CCR5 and ARG when compared to AIRmax animals. In late repair stages, the AIRmin strain presented a decreased (p<0.05) density of osteoclast and blood vessels than AIRmax, along lower RANKL and Catepk and higher PHEX and SOST mRNA expression, but the healing outcome at the endpoint was similar in AIRmin and AIRmax strains. When analyzed the effect of RR/SS Slc11a1 genotypes was evaluated in parallel with the influence AIRmin/AIRmax background, we initially observed that the AIRmax strain, associated with both RR and SS Slc11a1 genotypes, presented a more effective bone healing, characterized by increased (p<0.05) of bone volume and predominance of red fiber in analysis in contrast to AIRmin strains. AIRmaxRR presented increased (p<0.05) F4/80+ and decreased CD80+ e CD206+ cells count, while AIRmaxSS presented increased (p<0.05) GR1+, F4/80+ and CD80+ and decreased CD206+ cells. When the analysis was performed in order to address the influence Slc11a1 variants, AIRmaxSS strain presented a bone healing delay when compared to AIRmaxRR; characterized by decreased (p<0.05) of bone volume, trabecular number and red collagen fibers, increased (p<0.05) GR1+ and CD80+ and decreased F4/80+ and CD206+. Conversely, AIRminSS presented a more effective healing when compared with AIRminRR mice; characterized by increased (p<0.05) of bone volume, trabecular number/separation and red birefringence, increased GR1+ and decreased CD206+ cells count. In conclusion, while AIRmin and AIRmax strains presents a similar healing outcome at the endpoint, the early repair in AIRmin strain was associated with decreased presence of neutrophils and M1 macrophages, and increased M2 macrophages. Additionally, our while results showed that AIRmax inflammatory background was associated to a more effective bone healing process irrespective of the presence of RR/SS Slc11a1 genotypes, RR genotype favors the healing in AIRmax background and SS genotype was found to favor the healing in the AIRmin background.(AU)

O processo de reparo ósseo alveolar pode ser influenciado por vários fatores locais e sistêmicos, que incluem o sistema imunológico e os genes relacionados ao reparo. No entanto, o exato papel da resposta inflamatória do hospedeiro e genético background no processo de reparo ósseo ainda não está claro. Neste contexto, avaliamos a influência da inflamação no reparo óssea alveolar, em camundongos selecionadas geneticamente para uma resposta inflamatória aguda máxima (AIRmax) ou mínima (AIRmin), como também em camundongos AIR homozigoto para os alelos RR/SS do gene Slc11a1.Neste estudo foram utilizados camundongos machos e fêmeas (N=5/tempo/grupo), das linhagens selecionados para máxima e mínima (AIRmax e AIRmin) reação inflamatória, e também as sublinhagens AIRminRR, AIRminSS, AIRmaxRR e AIRmaxSS com idade aproximada de 8 semanas. Todos foram submetidos à extração do incisivo superior direito e avaliados nos períodos de 0, 3, 7, 14 e 21 dias pós extração, seguido pela análise tomografia computadorizada (CT), análise histomorfometria, análise de birrefringência, análise imuno-histoquímica e análise molecular (PCRArray). Inicialmente, nossos resultados demonstraram que a linhagem AIRmin, no período inicial, apresentou um aumento (p<0.05) no volume ósseo, nas regiões hiperdensas, na densidade de matriz óssea e osteoblastos, seguido pelo aumento (p<0.05) na expressão de BMP4, BMP7 e RUNX2 quando comparado a linhagem AIRmax. Camundongos AIRmin também apresentou uma menor contagem de células GR1+ e CD80+ e aumento da contagem de células F4/80+ e CD206+, em paralelo com aumento da expressão de mRNA de CX3CL1, CCL5, CCR5 e ARG quando comparado aos camundongos AIRmax. Nos períodos tardios, a linhagem AIRmin apresentou uma diminuição (p<0.05) na densidade de osteoclastos e vasos sanguíneos em comparação AIRmax, seguido por uma diminuição na expressão de mRNA de RANKL e Catepk e aumento de PHEX e SOST, mas o processo de reparo ósseo alveolar, no período final foi semelhante entres as linhagens AIRmin e AIRmax. Quando analisamos o efeito dos alelos RR/SS do gene Slc11a1 em paralelo com a influência do background AIRmin/AIRmax, nós inicialmente observamos que a linhagem AIRmax associada com ambos os alelos RR/SS do gene Slc11a1 apresentaram um processo de reparo mais efetivo, caracterizado pelo aumento (p<0.05) volume ósseo e predominância de fibras vermelhas em comparação com a linhagem AIRmin. Camundongos AIRmaxRR apresentaram aumento (p<0.05) na contagem de células F4/80+ e diminuição na contagem de células CD80+ e CD206+, enquanto, camundongos AIRmaxSS apresentou um aumento (p<0.05) na contagem de células GR1+, F4/80+, CD80+ e diminuição na contagem de células CD206+. Quando analisamos a influência dos alelos do gene Slc11a1, a linhagem AIRmaxSS apresentaram um atraso no reparo óssea quando comparado ao AIRmaxRR; caracterizado pela diminuição (p<0.05) do volume ósseo, número trabecular e fibras colágenas vermelhas, seguido pelo aumento (p<0.05) da contagem de células GR1+ e CD80+ e diminuição de células F4/80+ e CD206+. Por outro lado, camundongos AIRminSS apresentaram um reparo ósseo mais efetivo quando comparada com AIRminRR; caracterizada pelo aumento (p<0.05) do volume ósseo, número / separação trabecular e birrefringência das fibras colágenas no espectro vermelho, seguido pelo aumento da contagem de células GR1+ e diminuição das células CD206+. Diante disso, os nossos resultados demonstraram que as linhagens AIRmin e AIRmax apresentaram um processo de reparo ósseo alveolar semelhantes no período final do reparo, enquanto no reparo inicial a linhagem AIRmin estava associada com a diminuição de neutrófilos e macrófagos M1 e aumento dos macrófagos M2. Além disso, nossos resultados demonstraram que background AIRmax estava associado a um processo de reparo mais efetivo, independentemente da presença de genótipos RR/SS Slc11a1, o genótipo RR favorece o reparo no background AIRmax e o genótipo SS favoreceu a reparo no background AIRmin.(AU)

Association of the solute carrier family 11 member 1 gene polymorphisms with susceptibility to leprosy in a Brazilian sample

Natural resistance-associated macrophage protein 1/solute carrier family 11 member 1 gene (Nramp1/Slc11a1) is a gene that controls the susceptibility of inbred mice to intracellular pathogens. Polymorphisms in the human Slc11a1/Nramp1 gene have been associated with host susceptibility to leprosy. This study has evaluated nine polymorphisms of the Slc11a1/Nramp1 gene [(GT)n, 274C/T, 469+14G/C, 577-18G/A, 823C/T, 1029 C/T, 1465-85G/A, 1703G/A, and 1729+55del4] in 86 leprosy patients (67 and 19 patients had the multibacillary and the paucibacillary clinical forms of the disease, respectively), and 239 healthy controls matched by age, gender, and ethnicity. The frequency of allele 2 of the (GT)n polymorphism was higher in leprosy patients [p = 0.04, odds ratio (OR) = 1.49], whereas the frequency of allele 3 was higher in the control group (p = 0.03; OR = 0.66). Patients carrying the 274T allele (p = 0.04; OR = 1.49) and TT homozygosis (p = 0.02; OR = 2.46), such as the 469+14C allele (p = 0.03; OR = 1.53) of the 274C/T and 469+14G/C polymorphisms, respectively, were more frequent in the leprosy group. The leprosy and control groups had similar frequency of the 577-18G/A, 823C/T, 1029C/T, 1465-85G/A, 1703G/A, and 1729+55del4 polymorphisms. The 274C/T polymorphism in exon 3 and the 469+14G/C polymorphism in intron 4 were associated with susceptibility to leprosy, while the allele 2 and 3 of the (GT)n polymorphism in the promoter region were associated with susceptibility and protection to leprosy, respectively.

Differential expression of solute carrier family 11a member 1 and inducible nitric oxide synthase 2 in skin biopsies from leprosy patients.

Background: Leprosy is a chronic granulomatous infection caused by Mycobacterium leprae, an intracellular parasite that resides within macrophages and cannot be eliminated effectively. Solute carrier family 11a member 1 (Slc11a1) and inducible nitric oxide synthase (iNOS), both expressed in macrophages, play major roles in host defense against several intracellular pathogens. However, the roles of these molecules in natural infection with M. leprae remain unknown. Objective: We aimed to investigate the expression of Slc11a1 and iNOS in macrophages (CD68+ cells) infi ltrating skin lesions in leprosy. Methods: Skin biopsies from 48 Mexican patients of leprosy [(33 lepromatous (LL), 15 tuberculoid (TT)] and from 10 healthy controls, were subjected to immunohistochemistry to determine expression of CD68, Slc11a1 and iNOS. Results: We found a high expression of Slc11a1 and iNOS in most lepromatous leprosy samples. In tuberculoid leprosy samples, Slc11a1 expression was moderate or low, and that of iNOS was almost always low. In addition, Slc11a1 and iNOS expression levels were positively associated with bacillary loads in lepromatous leprosy lesions (P = 0.05). Conclusions: These observations suggest that M. leprae infection promotes the expression of Slc11a1 and iNOS in macrophages and that lepromatous leprosy can occur despite this response.

Influência dos alelos R e S do gene Slc11a1 na suscetibilidade à carcinogênese de pele induzida por 7, 12-dimetilbenzantraceno / Influence of R and S alleles of Slc11a1 gene in the susceptibility to skin cancer induced by 7 12-Dimethylbenz[a]anthracene

Camundongos AIRmax e AIRmin diferem na sensibilidade à carcinogênese, sendo os AIRmin mais sensíveis à carcinogênese de pele devido ao seu fundo genético e um polimorfismo no gene Ahr. Mais que isso, estas linhagens possuem um desequilíbrio de frequência dos alelos do gene Slc11a1. Para estudar a interação dos alelos de resistência (R) ou suscetibilidade (S) do gene Slc11a1 com os loci de resposta inflamatória aguda dos animais AIRmax e AIRmin, foram produzidas sublinhagens homozigotas para estes alelos:AIRmaxRR, AIRmaxSS, AIRminRR e AIRminSS. Nosso objetivo foi investigar a diferença de sensibilidade à carcinogênese de pele induzida por DMBA nestas sublinhagens. A incidência de câncer de pele foi de 7% nos animais AIRminRR e de 13% nos animais AIRminSS.Os animais AIRmaxSS não apresentaram câncer de pele, mas a incidência de câncer em órgãos internos foi 100% nesta sublinhagem.Esses dados mostraram que os camundongos AIRmaxSS tem maior suscetibilidade à carcinogênese, sugerindo que o alelo S, no fundo genético AIRmax, pode influenciar na suscetibilidade ao câncer.

Mice AIRmax and AIRmin differ on sensibility to carcinogenesis. AIRmin mice are significantly more sensitive to skin carcinogenesis than AIRmax mice due to the genetic background and the polymorphism of aryl hydrocarbon receptor (Ahr) gene. Furthermore,these mice have an imbalance of frequency of Slc11a1 gene alleles.To study the interaction of resistant (R) or susceptible (S) Slc11a1 alleles with acute inflammatory reaction loci found in AIRmax and AIRmin mice, homozygous sublines for these alleles were produced: AIRmaxRR, AIRmaxSS, AIRminRR and AIRminSS. The objective of this study was to investigate the difference in skin carcinogenesis sensibility induced by DMBA agent in these sublines.The incidence of skin cancer was 7% in AIRminRR mice and 13% in AIRminSS mice.AIRmaxRR and AIRmaxSS mice did not show skin cancer, but the incidence of internal organs cancer was 100% only in AIRmaxSS mice. These data showed that AIRmaxSS animals have higher susceptibility, suggesting that the S allele in the AIRmax background could influence susceptible to cancer.

SLC11A1 gene mechanism and its relationship with susceptibility to tuberculosis / 国际儿科学杂志

Tuberculosis is the leading cause of death due to infection from a single microbial agent. With indepth study of the pathogenesis of tuberculosis,it was found that solute carrier family 11 member I gene SLC11A1) was ignificantly associated with susceptibility to tuberculosis. SLC11A1 is located in phagocytic lysosome membrane,influencing the occurrence and development of tuberculosis by changing the intracellular environment. In this paper, the mechanism of SLC11A1 gene and its relationwith susceptibility to tuberculosis are reviewed.