Expresión génica de mediadores moleculares para potencial terapia celular en el síndrome gastrointestinal inducido por radiación / Genetic expression of molecular mediators for potential cell therapy in radiation-induced gastrointestinal syndrome

Antecedentes: Las células madres intestinales generan las distintas estirpes celulares a dicho nivel. Estas se regulan por interacciones entre el epitelio y las células del nicho celular anexo. Estas se pueden ver dañadas en tratamientos con radiación, generando el síndrome gastrointestinal inducido por radiación. Se ha visto que células madre mesenquimales (MSC) y macrófagos de médula ósea (BMM) tienen propiedades de regeneración tisular. Objetivos: Evaluar la expresión génica de IL-4, Wnt6, VEGF y bFGF, a partir de cultivos celulares primarios independientes de MSC derivadas de tejido adiposo y BMM de ratones C57BL/6, por medio de PCR en tiempo real (qRT-PCR). Diseño experimental: A partir de un análisis in silico, se confeccionaron primers para evaluar la expresión génica de las moléculas propuestas, en los cultivos primarios por medio de qRT-PCR y electroforesis. Resultados y proyecciones: IL-4 y Wnt6 no son expresadas en las muestras de BMM y MSC. VEGF y bFGF son expresadas por diferentes células, dando expresión diferenciada. A futuro, se deben evaluar las mismas estirpes celulares en un ambiente inflamatorio y su efecto en la expresión génica, en especial VEGF y bFGF. Limitaciones: El número de moléculas en estudio es limitado y la expresión se evalúo solo a nivel genético.

Background: Intestinal stem cell generates diferents cellular types in their niche. They're regulated by interactions between epithelium and niche's cells, and can be damaged by medical radiation treatments causing radiation-induced gastrointestinal syndrome. It has seen that mesenchymal stem cells (MSC) d and bone marrow-derived macrophages (BMM) have propierties of tissular regeneration. Objectives: Determinated genetic expression of IL-4, Wnt6, VEGF and bFGF, in primary cellular cultures of MSC derivated of adipose tissue and BMM of C57BL/6 mice, through real time PCR (qRT-PCR). Methods: By an in silico analysis, we created primers to evaluate the proposed molecules in the primary cellular cultives, with qRT-PCR and electrophoresis. Results and projections: IL-4 and Wnt6 were not expressed in the MSC and BMM samples. VEGF and bFGF were expressed by different cells, giving differential expression. In the future, the same samples should be analyzed in an inflammatory environment, especially VEGF and bFGF. Limitations: The number of molecules are limited and the expression of them is only in a genetic level.

IL-4-C-590T locus polymorphism and susceptibility to asthma in children: a meta-analysis

Abstract Objectives The study aimed to evaluate the link between the IL-4-C590T polymorphism and asthma susceptibility in children by meta-analysis. Sources The study collected all the case-control studies found in PubMed, Embase, CNKI, Wanfang, VIP, and other databases until September 2019. Stata v. 15.0 was used to conduct meta-analysis, calculate the combined OR and its 95% CI, and then conduct subgroup analysis. Summary of the findings Seven studies were included in the study, containing 860 cases and 810 controls. Relative to the C allele, the T allele at the IL-4-C590T locus was associated with susceptibility to asthma in children (OR = 1.45, 95% CI: 1.05-2.01). The results of ethnicity subgroup analysis showed that there was statistical significance, with OR = 1.61 (95% CI: 1.01-2.57) in the Asian population. In the dominant and recessive genetic models, the overall test and the Asian population subgroup analysis were statistically significant. In the homozygous model, there was statistical significance, but no statistical significance in heterozygous model. Conclusions The IL-4-C590T polymorphism was associated with asthma susceptibility, and T allele and TT genotype may increase the risk of asthma susceptibility in children, especially in the Asian population.

Effects of Interleukin-4 and Interleukin-12b Gene Polymorphisms on Hepatitis B Virus Vaccination

Abstract: Approximately 10% of individuals do not respond to hepatitis B virus (HBV) vaccination, i.e. non-responders (NRs). We aimed to investigate the association of interleukin (IL)-4 and IL-12B gene polymorphisms with responsiveness to the HBV vaccine in Korean infants. Among 300 healthy infants (9-12 month), SNPs for the IL-4 gene (rs2243250, rs2070874, and rs2227284) and for the IL-12B gene (rs3213094 and rs17860508) were compared between subgroups in terms of the response to HBV vaccination. The percentages of NRs (< 10 mIU/mL), low-titer responders (LRs, 10-100 mIU/mL), and high-titer responders (HRs, ≥ 100 mIU/mL) were 20.3%, 37.7% and 42.0%, respectively. No SNPs differed in frequency between NRs and responders or between LRs and HRs. We divided the subjects into two groups according to the time interval from the 3rd dose of HBV vaccination to Ab quantification: > 6 months from the 3rd dose (n = 87) and ≤ 6 months from the 3rd dose (n = 213). In the ≤ 6 month subjects, rs2243250C and rs2227284G were significantly frequent in the lower-titer individuals (NRs + LR) than HRs (40.1 vs. 25.9%, p = 0.014 and 45.1 vs. 33.0%, p = 0.018, respectively), and the rs2243250C and rs2227284G frequencies were significantly different among the three subgroups (13.2 vs. 26.9 vs. 25.9%, p = 0.040 and 15.5 vs. 29.6 vs. 33.0%, p = 0.038, respectively). In conclusion, those results suggest that IL-4 gene polymorphisms may play a role in the response to the HBV vaccine in Korean infants.

Association between interleukin-4 [IL-4], gene polymorphisms [C-589T, T+2979G, and C-33T] and migraine susceptibility in Iranian population: a case-control study

Background: Migraine is a chronic neurological disease characterized by recurrent moderate to severe headaches commonly in association with neuro-inflammation. Interleukin-4 [IL-4], an anti-inflammatory cytokine, plays an important role in modulating pain threshold and has an essential role in stimulation of pain receptors in the trigeminal nerve fibers

Aim of the study: The current study aimed to investigate the possible associations between IL-4 single nucleotide polymorphisms [SNPs] and susceptibility to migraine in Iranian patients

Patients and methods: In a prospective case-control study, we studied blood samples of 190 patients with migraine [migraineurs] and 200 healthy controls [HCs] for analysis of gene variants. Genotyping for the IL-4 SNPs: C-589T [rs2243250], T+2979G [rs2227284], and C-33T [rs2070874] were performed using PCR-RFLP. Statistical analysis was performed using the SPSS version 21.0 [SPSS, Chicago] and SNPStats version 1.14.0

Results and Conclusion: Among IL-4 SNPs, rs2243250 [TC genotype, OR= 0.25, 95% CI= 0.13-0.50, P = 0.001] and rs2227284 [TG and TT genotypes, OR= 0.44, 95% CI= 0.23-0.92, P = 0.029 and OR = 0.38, 95% CI= 0.18-0.79, P = 0.009 respectively] were significantly associated with migraine. No significant associations between IL-4 SNP rs2070874 [TC, TT and CC genotypes] and migraine were found. The most frequent genotypes in the migraineurs were CC in both SNPs rs2243250 [79%], and rs2070874 [71.5%], as well as GG for SNP rs2227284 [64%]. There was no statistically significant relationship between these SNPs and different subclasses [common, classic and complicated] of migraine. Our findings revealed that in IL-4 rs2243250 and rs2227284 genotypes and allele frequencies have a role in susceptibility to migraine in our population. Therefore, it is suggested that in addition to other factors, IL-4 genetic variations also play a pivotal role in the progress of migraine

Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine

The purpose of this study was to investigate the effects of porcine interleukin (IL)-2 and IL-4 genes on enhancing the immunogenicity of a porcine reproductive and respiratory syndrome virus ORF5 DNA vaccine in piglets. Eukaryotic expression plasmids pcDNA-ORF5, pcDNA-IL-2, and pcDNA-IL-4 were constructed and then expressed in Marc-145 cells. The effects of these genes were detected using an indirect immunofluorescent assay and reverse transcription polymerase chain reaction (RT-PCR). Characteristic fluorescence was observed at different times after pcDNA-ORF5 was expressed in the Marc-145 cells, and PCR products corresponding to ORF5, IL-2, and IL-4 genes were detected at 48 h. Based on these data, healthy piglets were injected intramuscularly with different combinations of the purified plasmids: pcDNA-ORF5 alone, pcDNA-ORF5 + pcDNA-IL-2, pcDNA-ORF5 + pcDNA-IL-4, and pcDNA-ORF5 + pcDNAIL-4 + pcDNA-IL-2. The ensuing humoral immune responses, percentages of CD4+ and CD8+ T lymphocytes, proliferation indices, and interferon-gamma expression were analyzed. Results revealed that the piglets co-immunized with pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2 plasmids developed significantly higher antibody titers and neutralizing antibody levels, had significantly increased levels of specific T lymphocyte proliferation, elevated percentages of CD4+ and CD8+ T lymphocytes, and significantly higher IFN-gamma production than the other inoculated pigs (p < 0.05).

Genetic polymorphisms and asthma: findings from a case - control study in the Madeira island population

BACKGROUND: Asthma is a complex disease influenced by multiple genetic and environmental factors. While Madeira has the highest prevalence of asthma in Portugal (14.6%), the effect of both genetic and environmental factors in this population has never been assessed. We categorized 98 asthma patients according to the Global Initiative for Asthma (GINA) guidelines, established their sensitization profile, and measured their forced expiratory volume in 1second (FEV1) and forced vital capacity (FVC) indexes. Selected single nucleotide polymorphisms (SNPs) were analysed as potential markers for asthma susceptibility and severity in the interleukin 4 (IL4), interleukin 13 (IL13), beta-2-adrenergic receptor (ADRB2), a disintegrin and metalloprotease 33 (ADAM33), gasdermin-like (GSDML) and the signal transducer and activator of transcription 6 (STAT6) genes comparatively to a population reference set. RESULTS: Although mites are the major source of allergic sensitization, no significant difference was found amongst asthma severity categories. IL4-590*CT/TT and IL4-RP2*253183/183183 were found to predict the risk (2-fold) and severity (3 to 4-fold) of asthma and were associated with a lower FEV1 index. ADRB2-c.16*AG is a risk factor (3.5-fold), while genotype GSDML-236*TT was protective (4-fold) for moderate-severe asthma. ADAM33-V4*C was associated to asthma and mild asthma by the transmission disequilibrium test (TDT). Finally, ADAM33-V4*CC and STAT6-21*TT were associated with higher sensitization (mean wheal size ≥10mm) to house dust (1.4-fold) and storage mite (7.8-fold). CONCLUSION: In Madeira, IL4-590C/T, IL4-RP2 253/183, GSDML-236C/T and ADAM33-V4C/G SNPs are important risk factors for asthma susceptibility and severity, with implications for asthma healthcare management.

Cloning and expression of Bos indicus interleukin-4 in mammalian cells.

Dendritic cells (DC) which are located at the interface of innate and adaptive immunity are targets of infection by many RNA and DNA viruses. Advances in the ex vivo generation of monocyte derived non proliferating dendritic cells have been used for clinical application like immunotherapy. IL-4 cytokine plays essential role in the maturation and generation of DCs. Bos indicus interleukin 4 (boIL-4) 408 bp was amplified from PBMC’s and cloned in pBSIIKS+ vector. The sequence analysis showed N terminal 69 bp signal sequence and one N-glycosylation site. The phylogenetic tree analysis showed that Bos indicus IL-4 is closely related to the ruminant IL-4 and least sharing of genetic line of human and mouse IL-4. The recombinant boIL-4 protein was expressed in CHO cells which secreted a 16 kDa protein which was confirmed by SDS PAGE and western blotting. The rec-boIL-4 protein proliferated the bovine PBMC’s, decreased production of nitric oxide in antigen stimulated macrophages, and phagocytosed the micro particles confirming its activity on dendritic cells.

Association of interleukin-4 and interleukin-17F polymorphisms in periodontitis in Dravidian ethnicity.

BACKGROUND: Complex network of pro and anti-inflammatory cytokines are known to act in inflamed periodontal tissue. This study explores the distribution of interleukin (IL)-4 (+33 C/T) and IL-17F (7383A/G, 7488A/G) gene polymorphism in chronic and aggressive periodontitis subjects of Dravidian ethnicity. MATERIALS AND METHODS: This case control study consisted of 124 periodontitis individuals comprising of 63 chronic and 61 aggressive periodontitis subjects as cases, and control group consisted of 101 healthy subjects. All subjects were genotyped for IL-4 + 33C/T, IL-17F 7383A/G, 7488A/G by polymerase chain reaction amplification followed by TaqMan assay for IL-4 + 33C/T, restriction enzyme digestion and gel electrophoresis for IL-17F 7383A/G and sequencing for IL-17F 7488A/G. RESULTS: IL-4 + 33C/T was significantly associated with periodontitis (P < 0.05) at both allelic and genotypic level. In subgroup analysis also significant difference (P < 0.05) in allelic distribution between aggressive periodontitis and control group for loci IL-4 + 33C/T was noted. However, there was a lack of association between IL-17F 7383A/G and IL-17F 7488A/G with periodontitis and its sub-groups at both allelic and genotypic levels. CONCLUSIONS: In Malayalam speaking Dravidian population IL-4 + 33C/T loci appears to be an important risk factor for periodontal disease with a leaning towards aggressive periodontitis. The association between IL-17F at 7383A/G and 7488A/G loci with either chronic or an aggressive periodontitis could not be ascertained.

IFN-gamma +874 and IL-4 -590 polymorphisms and asthma susceptibility in North West of Iran

Recent studies have documented an association between some cytokines' gene polymorphisms and chronic inflammation of the respiratory tract which leads to asthma susceptibility. This study was conducted to investigate if there were any differences in IFN-gamma +874 [A/T] and IL-4 -590 [C/T] single nucleotide variations in asthmatic patients compared to normal controls among West Azerbaijani population. IFN-gamma +874 [A/T] and IL-4 -590 [C/T] polymorphisms were amplified by ASO-PCR and RFLP-PCR from genomic DNA of 173 individuals including 64 asthmatic patients and 109 control subjects from West Azerbaijani population. The allele or genotype frequencies of IFN-gamma+874 A/T in patients were not different from those of controls [p>0.05]. The differences between allelic or genotypic frequencies of IL-4 -590 C/T in patients and controls were not statistically significant [p>0.05]. These findings showed that IL-4 -590 [C/T] and IFN-gamma+874 [A/T] polymorphisms were not associated with asthma susceptibility

Do IL-4 intron 3 VNTR and IL-6 (-174) G/C variants reflect ethnic variation? A comparative study between the global and North Indian populations.

Variations in the production and activity of cytokines have been reported by several investigators which influence the susceptibility and/or resistance to various infectious agents and cancer. Differences in the cytokine production between individuals are often caused by single nucleotide polymorphism (SNP) in the promoter or coding regions of cytokine genes. Although the SNP cytokine gene variations are basically mutations, they are designated as polymorphisms, because these changes do not modify the alleles to rare or abnormal variants. The two important cytokine genes IL-4 and IL-6 of 343 unrelated healthy individuals from North India were compared with the published polymorphism of other populations. It was seen that our population differs from South Indian population as well as from other Caucasian populations except, Taiwanese population at IL-4 locus and Spanish and Polish population at the IL-6 gene locus. This study may be helpful for predicting clinical outcome of various infectious and immunoregulated disorders as well as explore for risk alleles for various cancers.

Relation of Interleukine-4 [C-589T] and Interferon-Gamma [A+874T] allele polymorphism with Asthma

Asthma is a common respiratory disease caused by acute and chronic bronchial inflammation. Clinical manifestations of the disease are closely related to genetics. IL-4 is a cytokine of TH2 lymphocytes, polymorphism in prompter region, C-589T, is associated with IL-4 production, while IFN-gamma, is a cytokine of TH1, and A+874T polymorphism in interon 1 of IFN-gamma is associated with it.s production and release. Cytokine gene polymorphisms could influence pathogenesis of asthma with TH1/THh2 ratio, being of great importance. 81 unrelated asthmatic patients were selected according to ATS characteristics and separated into two groups of controlled and uncontrolled asthma. 80 normal subjects were chosen as control group. After collection of peripheral blood and DNA extraction, PCR-RFLP method was used for genotyping of IL-4, -589 position. For evaluation of polymorphism in +874 position of IFN-gamma ARMS-PCR method was used. Distribution of frequency of IFN-gamma [A+874T] and IL-4[C-589T] polymorphisms didn.t show any statistically significant difference between two patient groups and healthy control group [p >/= 0.05]. There was neither any significant difference [p >/= 0.05] among other parameters. Studies in field of cytokine polymorphism have had variable results. So many studies have mentioned a relationship between cytokine gene polymorphism and susceptibility and/or severity of asthma and some studies have shown that there is no association between these factors we believe that there may exist factors different from IL-4 and IFN-gamma polymorphism which coner the effects of these genetic vaciants in pathogenesis and severity of asthma

No association between an interleukin 4 gene promoter (-589) polymorphism and Graves' disease in Thai patients.

Cytokines play a key role in the regulation of immune and inflammatory responses and therefore are potential candidate genes for autoimmune thyroid disease. Polymorphisms in cytokine genes may effect gene transcription, causing individual variations in cytokine production. Several investigators have linked the interleukin-4 (IL-4) gene and autoimmune disease. The present population-based study was to investigate the polymorphisms of IL-4 gene promoter (-589C/T) in GD patients compared with a control group and determine the association with GD in a Thai population. The subjects included 137 GD patients and 137 healthy control subjects with similar ethnic and geographic backgrounds. The IL-4 gene polymorphism at position -589 in the promoter was analyzed using the PCR-RFLP. The protective effect of the -589T allele as suggested by Hunt et al in a Caucasian population was not observed in the present study. The -589T allele frequencies were similar between patients and control subjects (69% vs 69.3%) suggesting that this polymorphism can not be used as a genetic marker for GD susceptibility in Thais.

Polymorphisms of IL-4, IL-4R alpha, and AICDA genes in adult allergic asthma / 华中科技大学学报（医学）（英德文版）

The relationship between 3 polymorphisms sites [interleulin-4 (IL-4), IL-4 receptor (IL-4R) alpha chain and activation-induced cytidine deaminase (AICDA)] and adult allergic asthma in China was studied. By using case-control method, DNA and clinical data were obtained from allergic asthmatic patients and compared with those in the control subjects. The subjects were genotyped for the IL-4 C-589T promoter polymorphism, the IL-4R alpha chain Q576R and the AICDA C8408T by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The results showed that the IL-4 C-589T was not associated with adult allergic asthma in China. However, the IL-4R alpha chain 576R/R and AICDA 8408T/T frequency was significantly increased in allergic asthma group as compared with that in the control group [odd ratio (OR) = 3.797 and 9.127, respectively; P < 0.01)] and was correlated with the increased plasma total IgE. These data suggested that the IL-4R alpha chain 576R/R and AICDA 8408T/T genotypes confer genetic susceptibility to adult allergic asthma in China.