Research/Original Article- Immunology

The frequency of human leukocyte antigen-DRB1 alleles, using sequence-based genotyping in 68 parents-child trios study in Iranian subjects

Zahra Golchehre, Majid Kabuli, Ahmad Salimzadeh, Mohsen Akhiani, Karim Faraji, Somayeh Ahmadlou, David Sayer, Arash Salmaninejad, Daniz Kooshavar, Mohammad Keramatipour

Archives of Medical Laboratory Sciences, Vol. 2 No. 2 (2016), 16 Shahrivar 2016,

Background: The human leukocyte antigen-DRB1 (HLA-DRB1) locus is one of the most polymorphic human loci and has a crucial role in the immune system. Assessing the allelic frequencies of HLA-DRB1 locus would be a fundamental factor in defining the origin of populations, relationships with other populations, disease association studies and the constitution of unrelated bone marrow donor registries. In the current study HLA-DRB1 alleles and their frequencies are determined in a family-based study by DNA sequencing-based typing high-resolution (2 field) level of typing.
Materials and Methods: Genomic DNA from 3 members of 68 unrelated families (a total of 204 individuals) was extracted. Exon 2 of DRB1 gene was amplified and performed useing AssignTM SBT v4.7 sequence analysis software.

Results: We had DRB1*11:04 with frequency of 0.0931, DRB1*03:01 with 0.0882, DRB1*11:01 with 0.0735, DRB1*13:01 with 0.071 and also alleles DRB1*08:03, DRB1*13:42, DRB1*14:04 and DRB1*14:07 with frequency of 0.0024.

Conclusion: A total of 34 different alleles were found in the study subjects with DRB1*11:04, DRB1*03:01, DRB1*11:01 being the most frequent alleles respectively.




Effects of black seed (Nigella Sativa) on type 2 cytokines gene expression and mucus production in the airways of asthmatic mice

Ebrahim Azimi, Seyyede Masoume Athari, Faride Afshari, Asie Eftekhari, Seyyed Shamsadin Athari

Archives of Medical Laboratory Sciences, Vol. 2 No. 2 (2016), 16 Shahrivar 2016,

Background: Black Seed (BS) is used in traditional medicine as a therapy for a variety of diseases including allergic asthma.
Materials and Methods: In the present study, anti-inflammatory and immunomodulatory effects of BS on cytokine gene expression, lung airway eosinophilia and goblet cell hyperplasia were examined in a mouse mice model of allergic asthma. Groups of 6-week-old female BALB/c mice were sensitized by intraperitoneal injections of OVA plus alum on days 1 and 14. On days 24, 26, 28 and 30, the mice were exposed to OVA in saline for 30 min with nebulizer. Similar experiments were conducted with mice receiving saline as a negative control.
Results: The mouse allergic asthma model received BS by food on days 23, 25, 27 and 29. Then, the percentage of inflammatory cells as well as mRNA expression levels of interleukin (IL)-4, IL-5, IL-13 and mucin (MUC5a) genes were survived in Broncho-alveolar lavage fluid (BALF). Furthermore, we attempted to examine histopathological examination of the lung. Mice receiving BS showed a significant decrease in the number of eosinophils, and a potential inhibitory effect on mRNA expression levels of Th2-driven immune response cytokines and mucin, resulting in decreased production of interleukin and mucin in allergic asthma.
Conclusion: Our findings suggested that BS has an anti-inflammatory and immunomodulatory effect during the allergic response in the lung, and can be a promising treatment for allergic asthma in humans.

The effect of Migri-Heal® on nitric oxide production in an in vitro inflammatory model of primary microglial cells

Mahmoud Hassani, Farzaneh Sabouni, Solaleh Emamgholipour, Mohammad Hessam Rafiee, Mohammad-Sadegh Fallah, Shah-Sanam Abbasi, Mohammad Ansari

Archives of Medical Laboratory Sciences, Vol. 2 No. 2 (2016), 16 Shahrivar 2016,

Background: Recently, much attention has been directed towards considering activated microgelial cells as putative targets for treatment of neurological disorders. MigriHeal® as a novel herbal remedy was introduced for the treatment of migraine headaches. The previous researches has shown that MigriHeal® extracts can decrease NO in an in vitro inflammatory model. The aim of this study was to investigate the effect of MigriHeal® on NO generation from LPS- stimulated microglia cells.

Materials and Methods: Neonatal rat primary microglial cells were isolated from the mixed glial cultures and the purity of the cultures was determined by immunocytochemistry. Microglial cells were pretreated with Migri-Heal® and activated by 1μg/ml LPS. Subsequently, NO levels in the culture supernatants were measured by a griess reaction. Our results showed that Migri-Heal® 50μg/ml significantly reduced NO level in inflamed microglia in a dose-dependent manner. Results: The results showed that different concentrations of Migri-Heal® had no prominent effect on cell viability in presence of LPS as compared with the control group. In addition, the pretreatment of microglia cells with Migri-Heal® can prevent from a morphological changes of the cells into the round and phagocytic shape. Conclusion: Our study demonstrated that MigriHeal® might have NO scavenging properties. Integrative studies are warranted to uncover the novel pharmacological insights of this herbal remedy as an putative therapeutic approach against diseases - associated with inflammation.

Autophagy knock down: as a booster for the replication of viruses in cell culture

Mahdieh Talebi, Taravat Bamdad, Ehsan Arefian, Habib Ahmadi, Hesam Karimi, Hamzeh Choobin

Archives of Medical Laboratory Sciences, Vol. 2 No. 2 (2016), 16 Shahrivar 2016,

Background: Autophagy suppression recently has been known to have a remarkable effect for cellular adjustment and viability in the final stages of cancer. On the other hand, autophagy has the potential effect in preventing many viruses from replication. Beclin1 is the most substantial constituent in autophagy apparatus regulation. This study was intended to investigate the beclin1 siRNA knockdown effect on the extent of activity of the oncolytic vesicular stomatitis virus (VSV) as a model in cell culture.

Materials and Methods: In the current study, the cancer cell line , HeLa (cervical squamous cancer cell line ) was infected by VSV, followed by beclin1 siRNA vector transfection. The potential change in the expressions of gene beclin1 in transfected cells, as well as untransfected ones were examined by real time PCR, and also the titer of viruses was compared in cells with and without transfection.

Results: The results revealed that the amount of putative gene beclin1 expression in HeLa cells decreased greatly due to siRNA suppressive impact, and also the sensitivity of the cells to VSV oncolytic effect increased upon decrease in beclin1gene expression.

Conclusion: It seems that autophagy suppression by using siRNA with VSV is a substantial aid for increase in virus titer in cancer cell lines.

Comparison of the Prevalence of Human Papillomaviruses among Fertile and Infertile Women in Mashhad, Northeast of Iran

Aida Javanbakht, Zahra Meshkat, Nayereh Khadem Ghaebi, Mojtaba Meshkat, Sedigheh Ayati Afin, Narges Javan, Sina Rostami

Archives of Medical Laboratory Sciences, Vol. 2 No. 2 (2016), 16 Shahrivar 2016,

Background: Human papillomaviruses (HPVs) are the most common viruses which can be sexually transmitted. They can cause different malignancies in asymptomatic women. The association of HPVs with infertility among men and women is controversial. In the current study, the authors compared the frequency of HPVs in fertile and infertile women in the city of Mashhad.
Materials and Methods: In the present case-control study, cervical and vaginal smears were collected from infertile and fertile women. HPVs were detected by polymerase chain reaction. Data was analyzed by SPSS v.20 and P-value <0.05 was considered statistically significant.
Results: In the current study, 115 infertile women with the mean age of 30.5±5.6 years and 60 fertile women with the mean age of 32.6±9.3 years were included (p=0.07). Among women who were infertile (cases), 121 (52.6%) of 230 smears were positive, while in control group (who were fertile), 50 (41.7%) of 120 smears were positive (p=0.052).
Conclusion: Frequency of HPV in both groups was high, which could be due to lack of routine HPV vaccination. HPV can cause placenta abnormality, our infertile women had multiple abortion history and history of abortion had significant differences among infertile and control group. The frequency of HPV had no significant differences between the infertile and control groups.

Short Communication

 Background: Alpha-thalassemia syndrome includes a group of hereditary anemia in which expression of alpha globin chains is decreased or absent. Impaired RBC in patients with thalassemia causes vessel involvement and endothelial cell vessel disturbance. Vascular Endothelial Growth Factor (VEGF) is the most important regulator for endothelial cell proliferation. So, the aim of this study is to compare the serum VEGF levels in patients with alpha thalassemia and normal control group.

Materials and Methods: This case-control study was conducted on 17 patients with alpha thalassemia and 40 healthy people. Serum VEGF levels were measured by enzyme-linked immune sorbent assay (ELISA) kit. Then statistical analysis of results were performed using SPSS 16, value of P <0.05 was considered statistically significant.

Results: Mean serum VEGF levels in case and control groups were 2294.19±1552.39 and 598.09±988.17pg/ml, respectively. Serum VEGF levels were higher in patients with alpha thalassemia (P <0.01). There was no significant correlation between serum VEGF levels and Hemoglobin. (P= 0.73).

Conclusion: Our study revealed that patients with alpha thalassemia have elevated levels of serum VEGF than normal control group. Further studies with larger sample size are recommended to confirm these observations.