Immunogenetic study of parvovirus B19 infection and IL12 gene polymorphism in women with recurrent miscarriage

Abstract

Viral infections during pregnancy have been associated with adverse pregnancy outcomes and birth defects in the offspring. Viruses rarely cross the placental barrier, but when the virus does reach the fetus, it can result in severe birth defects such as microcephaly or even fetal death. Human parvovirus (B19) is thought to cause severe complications is spontaneous abortion. Normal pregnancy is related to the successful transition from type 1 cellular immunity to type 2 cellular immunity. Interleukin (IL)-12 induced the production of type 2 cytokines. These experiments were conducted to study the effect of human B19 virus on the pregnant women and cause abortion. This case control study was done for 100 patients including different ages that range from 18-42 age that sever recurrent miscarriage. Also, the study include 50 apparently healthy control (AHC) this age were similar with the patients age. They were collected from different general hospitals in hilla; Baghdad as well as Mid-Euphrates Governorates of Iraq, during the period from February 2021 to September 2021. Endometrium; Cervical swabs; fetal fluids swabs as well as Blood specimens were collected and processed to extract viral genome and total DNA gene for screened human B19 virus by using PCR and IL-12(rs 3212227) polymorphism by PCR and sequencing. In addition, estimation serum IL-12 concentration by enzyme-linked Immunosorbent assay (ELISA). Out of 150 Endometrium; Cervical swabs; fetal fluids swab as well as Blood specimens involved in this study 48 % (72 out of 150) were found to have a viral infection with abortion & RPL. In women with RPL, the most commonly affected age stratum infected with DNA – B19 was (30-39 years) which constituted 48% (12 out of 25 cases), while the age stratum (17-29 years) was constituted 32% (8 out of 25 cases), followed by 20% (5 out of 25) in age stratum (40 – 49 years).Statistical comparison of these B19 in the Patients with Women With RPL according to age stratum revealed significant differences (p< 0.05). The percentage of a single band (162 bp) of the target sequence of IL-12 rs3212227 gene. The positive result, according to PCR amplification of a single band (162 bp) of IL-12 rs3212227 gene in women patients with RPL and AHC were 36.7% (55 of 150 cases) and 20% (5 of 50 cases), respectively. While the negative results were in women patients with RPL and AHC were 63.3% (95 of 150 cases) and 90% (45 of 50 cases), respectively. Two types of mutation in IL-12 rs3212227 gene was found transversion and transition. The frequency of transversion mutation (A\C; A\T; T\G; T\A and T\C) more than the transition mutation (A\G). The mean of serum IL-12 concentration for AHC and women patients with RPL groups were 8.00 ±0.31pg./ml and 14.00 ±0.59 pg./ml, respectively. Statistically, significant difference (p<0.05) was found on comparing the mean of serum IL-12 concentration among these study groups.