Method Article - (2023) Volume 14, Issue 2
Background: One of the most prevalent chronic diseases that affect children is type 1 diabetes. Transaction between hereditary weakness and natural variables is remembered to give the central component to the sickness. Aside from the Significant Histocompatibility locus which is the fundamental supporter of chance vulnerability, in excess of 40 loci are perceived. The CTLA-4 is one of these, but the data from the literature are disputed. Our research sought to determine whether CTLA4 49 A/G served as a risk factor for the onset of type 1diabetes in a group of Egyptian families.
Subjects and methods: This is a case-control study with 88 Egyptian families that have at least one index case younger than 18 years old. The control group consisted of 369 healthy, unrelated individuals who did not have a diabetes or autoimmune disease family history.
738 samples, including 369 controls and 88 families (88 patients, 125 siblings, and 156 parents), were used to test for CTLA4 49 A/G using PCRRFLP.
Results: The average age of onset ranged from 6 days to 12.5 years, with a median of 5 years and a mean of 5.3 3.6. 51 cases presented with classic symptoms, while 37 cases presented with diabetic ketoacidosis. Twentytwo of the cases had a history of viral infection or exanthematous disease, and four of them had autoimmune diseases that were associated with it. CTLA4 +49 A/G genotype or allele frequencies were not found to be significantly different in any of the groups. Neither the age of onset nor the mode of presentation were correlated with the various genotypes.
Conclusions: In our cohort, the CTLA4 49 A/G polymorphism was not identified as a risk factor. Due to the low incidence of autoimmune diseases, this may be the case. This is the first study involving families to our best knowledge. In order to prevent other autoimmune diseases from interfering with data analysis and making people more likely to develop type 1diabetes, we recommend that all studies on risk factors for the disease include thorough examination of those conditions.
Type 1 diabetes; CTLA-4 Risk susceptibility
One of the most prevalent chronic diseases that affect children is type 1 diabetes; Its prevalence varies greatly from population to population.
There have been reports of a prevalence rate in Egypt ranging from 0.7 to 1.9. People who are predisposed genetically are more likely to develop type 1 diabetes; it is believed that the primary cause of the disease is the interaction between environmental and genetic susceptibility.
Within a given environmental context, alleles or genetic variants associated with type 1 diabetes can either make a person more susceptible to the disease or protect them from it. The age at which type 1 diabetes first appears is determined by the genetic makeup's balance of susceptibility and protection alleles. The risk for affected individuals' siblings is roughly 15 times higher than the risk for the general population. The posterity of a mother with type 1 diabetes mellitus is 4-10 times while that for posterity of a diabetic dad is 15-22 times.The orientation predisposition in transmission rate has not been completely made sense of [1-3]. The major histocompatibility complex (IDDM1) is the primary genetic marker for type 1 diabetes susceptibility. However, despite their higher risk, HLA-identical siblings in diabetic families rarely develop diabetes. This brought attention to additional potential genetic loci, and more than 40 DDM loci are now recognized [18]. The cytotoxic T lymphocyte antigen-4 (CTLA-4) that is associated with insulin-dependent diabetes mellitus 12 (IDDM 12) is one of these. An amino acid substitution (Thr/Ala) occurs as a result of an A/G polymorphism in CTLA-4's first exon [4]. Type 1 diabetes, autoimmune thyroiditis, systemic lupus erythematosus, celiac disease, and biliary cirrhosis have all been linked to the presence of an alanine at CTLA-4's codon 17. However, there is a lot of debate about how it affects type 1 diabetes. On the one hand, it is claimed that the effect of IDDM12 is unrelated to any other genetic markers of type 1 diabetes. On the opposite side it is professed to be powerless or even sketchy. There has been evidence of ethnic diversity, with some populations experiencing strong effects and others experiencing weak transmission [5].
It is essential to investigate the primary loci that contribute to the susceptibility to or protection from type 1 diabetes in each community due to the potential ethnic variations and various environmental factors in each society; It is impossible to apply the findings of one population study to other populations.
The HLA region has been the primary focus of research to date on the genetic background of type 1 diabetes in Egypt, including ongoing research by the research team. The role of IDDM12 in type 1 diabetes risk has recently been the subject of two Egyptian studies; Families were not included in either of these case-control studies [6].
Subjects
88 Egyptian families with at least one index diabetic child or adolescent attended the study cohort's outpatient clinic at Cairo University Children Hospital's Diabetes Endocrinology and Metabolism Pediatrics Unit (DEMPU). 369 samples, including 88 families (88 patients, 125 siblings, and 156 parents) and 369 controls, were used to examine CTLA-4. The analysis did not include seven siblings with diabetes from six families; only the 118 siblings who were not diabetic were looked at [7].
With a mean age of 8.64 and a median age of 9 years, patients' ages at the time of the study ranged from 1 to 18 years. 53 of our patients were female, while 35 of our patients were male. The donors for the controls came from blood banks. They were unrelated, healthy individuals who did not have a family history of diabetes or other autoimmune diseases. The selection of grown-ups was to ensure that they wouldn't foster adolescent diabetes and to keep away from moral issues of involving solid youngsters as controls [8].
The salting out method was used to extract genomic DNA from EDTA peripheral blood samples for CTLA-4 +49 A/G
DNA analysis and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) testing. The PCR-RFLP protocol was used for the identification of the loci. Pharmacia biotech synthesized oligonucleotide primers. A 25 l PCR reaction with 100 ng of genomic DNA, 25 pmol of each primer (forward primer 5′- CCACGGCTTCCTTTCTCGTA-3′ and reverse primer 5′- AGTCTCACTCACCTTTGCAG-3′), 100 M of each dNTP, 1.5 mM of MgCl2, and 1.2 U of Taq polymerase (Promega, Madison) was used to amplify the CTLA-4 gene [9]. Initial PCR conditions included two minutes of denaturation at 95 °C, 40 cycles at 94 °C for 30 seconds, 50 °C for 45 seconds, and 72 °C for 30 seconds, and a final extension step at 72 °C for ten minutes. As a result, a 327-bp fragment is produced [10].
A 20-l mix containing 10 l of the PCR product, 10 U BbvI (Fermentas), and 1 X buffer was used to digest the product. For 15 minutes, the mixture was incubated at 65 °C. On 2% agarose, the digested products were separated. With a single band of 327 bp in the wild type (AA) and two bands of 244 bp and 83 bp in the G allele, there was no digestion (Fig. 1) in comparison to the Thermo-Fermentas 100 base pair ladder [11].
The onset age ranged from 6 days to 12.5 years, with a mean of 5 years and a median of 5 years. The range was 5.3 3.6 years. In 17 cases, the onset was less than a year old; Eight of them are less than six months old. The method of show was exemplary side effects in 51 and diabetic ketoacidosis (DKA) in 37 cases. Twenty-two of the cases had a viral infection or exanthematous disease before they got diabetes, and four of them had autoimmune diseases like Hashimoto's thyroiditis, rheumatoid arthritis, rheumatic heart disease, or Crohn's disease. 45 cases reported the early introduction of cereals or cow's milk, while the other cases exclusively breastfed for the first six months [12].
The CTLA-4 +49 A/G genotype and allele recurrence among the various gatherings are introduced in Table 1 and Fig. 1. There was no statistically significant difference in the distribution of CTLA-4 genotypes and alleles among patients and controls. AA versus AG + GG comparisons between patients and controls, non-diabetic siblings, and parents were also insignificant (p = 0.60, 0.49, and 0.68, respectively). Neither the manner of presentation nor the age at which symptoms first appeared was correlated with the various genotypes.
Currently, type 1 diabetes mellitus is a lifelong condition that requires a difficult treatment plan that only partially prevents acute and chronic complications. A better understanding of the genetics of type 1 diabetes mellitus in our community would make it easier to define the disease and identify people at risk for diabetes and its complications. We looked at 88 families with one or more children who had type 1 diabetes in this study. Female predominance was found in our study, which is consistent with the hypothesis that female predominance is prevalent in low-incidence regions, particularly among populations of non-European ancestry. This was refuted by a more recent study that found female predominance only in autoimmune diseases and an equal incidence for both sexes. Due to the fact that only four of our patients also have an autoimmune disease, this explanation does not apply to our situation.
According to our research, the early consumption of cereals or cow's milk did not influence the onset of type 1 diabetes. This is consistent with two studies—one from Iran and one from Finland. In the previous study, it was even claimed that cow's milk consumption at an early age decreased the likelihood of developing type 1 diabetes before the age of 8; an effect that was gone by the time she was 15 years old. In the latter, it was claimed that breastfeeding for a longer period of time is associated with protection, while breastfeeding for a shorter period of time is associated with an increased risk of type 1 diabetes. In Saudi Arabia, neither the early introduction of cereals nor cow's milk was found to have any effect. A history of exanthematous disease or viral infection was not linked to an increased risk of type 1 diabetes in our study. Cherian et al.'s findings are in line with this who investigated the influence of environmental factors on the likelihood of developing type 1 diabetes in Saudi Arabia's Eastern Province. Be that as it may, sporadic immunization and disease in the earlier a half year were accounted for as a trigger for the beginning of type 1 diabetes.
None
None
Indexed at, Google Scholar, Crossref
Indexed at, Google Scholar, Crossref
Indexed at, Google Scholar, Crossref
Indexed at, Google Scholar, Crossref
Indexed at, Google Scholar, Crossref
Indexed at, Google Scholar, Crossref
Received: 25-Jan-2023, Manuscript No. jdm-23-22187; Editor assigned: 28-Jan-2023, Pre QC No. jdm-23-22187(PQ); Reviewed: 11-Feb-2023, QC No. jdm-23-22187; Revised: 21-Feb-2023, Manuscript No. jdm-23-22187(R); Published: 28-Feb-2023, DOI: 10.35248/2155-6156.1000978
Copyright: © 2023 Kamele A. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
