Human Antibodies - Volume 24, issue 3-4

Authors: Basyreva, Liliya Yu | Brodsky, Ilya B. | Gusev, Alexander A. | Zhapparova, Olga N. | Mikhalchik, Elena V. | Gusev, Sergey A. | Shor, Dana Ben-Ami | Dahan, Shani | Blank, Miri | Shoenfeld, Yehuda

Article Type: Research Article

Abstract: INTRODUCTION: Intravenous immunoglobulin (IVIG) has been widely used to treat various conditions, including inflammatory and autoimmune diseases. IVIG has been shown to have a direct influence on neutrophils, eosinophils and lymphocytes. However, many aspects IVIG's effect on neutrophils activation still remain unknown. OBJECTIVE: To evaluate the effect of IVIG on PMA-induced activation of neutrophils, with and without priming with TNF-α, in a series of in vitro experiments performed on whole blood. RESULTS: Our data coincided with well-known literature indicating that the effect of phorbol 12-myristate 13-acetate (PMA) on human leukocytes includes activation …of neutrophils, monocytes and eosinophils, increase of chemiluminescence (CL) and induction of netosis, resulting in assembly of traps. In presence of IVIG (10 mg/mL), CL was reduced by 25% in response to PMA compared to PMA-induced leukocyte activation without IVIG. Decreasing IVIG concentration to 1 mg/mL and below did not inhibit PMA-induced activation of CL. PMA-induced activation after TNF-α priming resulted in approximately 50% increase of amplitude of CL response to PMA. Moreover, maximum CL was reached by minute 5, which was more rapid than in the absence of TNF-α-priming (in this case maximum CL was reached by minute 15). The IVIG concentrations did not affect morphological changes of leukocytes after sequential addition of TNF-α and PMA. IVIG had no effect on leukocyte content and on PMA-induced CL of primed leukocytes. Addition of IVIG under TNF-α priming significantly increased the number of traps in the smears in response to PMA activation. Of note, such an increase in the number of traps was depended on the IVIG concentration in plasma. CONCLUSION: In conclusion, we suggest that IVIG is able to reduce the degradation of traps under priming with TNF-α. Moreover, IVIG might switch the activation of primed leukocytes to netosis. Show more

Keywords: Intravenous Immunoglobulin (IVIG), autoimmunity, leukocytes, TNF-α, priming, neutrophils

DOI: 10.3233/HAB-160293

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 39-44, 2016

Authors: Barabas, Arpad Z. | Cole, Chad D. | Graeff, Richard M. | Morcol, Tulin | Lafreniere, Rene

Article Type: Research Article

Abstract: Objectives were to: 1) induce a lytic IgG antibody (ab) response (via the so called `third vaccination method') against CD38 antigen (ag) residing on the extra-cellular domain of multiple myeloma (MM) cells in recipient rabbits, by combining the CD38 ag with donor-derived anti-CD38 ag lytic IgG ab into an immune complex (IC); and 2) determine whether abs produced would cause complement-mediated lysis (in vitro ) of human MM cells containing CD38 ag. The vaccine was created in a two-step process. First, ab (rabbit anti-CD38 ag IgG ab) was raised in donor rabbits by injections of low molecular weight soluble CD38 …ag in Freund's complete adjuvant (FCA) and aqueous solution. Second, transfer of pathogenic lytic IgG ab response into recipient rabbits was achieved by injections of ICs composed of CD38 ag and homologous anti-CD38 ag IgG ab. Consequently, recipient rabbits produced the same ab with the same specificity against the target ag as was present in the inoculum, namely agglutinating, precipitating and lytic (as demonstrated in vitro ). In an in vitro study, in the presence of complement, donor and recipient rabbits' immune sera caused lysis of CD38 ag associated human MM cells. The most effective lytic ab response causing sera were those from donor rabbits injected with CD38 ag in FCA and those from rabbits injected with ICs, especially when they were administered in adjuvants. These results provided proof of concept that the third vaccination method has good potential as a stand-alone and efficacious method of controlling cancer. Show more

Keywords: Cancer, modified vaccination technique, prophylactic, therapeutic

DOI: 10.3233/HAB-160294

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 45-51, 2016

Authors: Barabas, Arpad Z. | Cole, Chad D. | Graeff, Richard M. | Kovacs, Zoltan B. | Lafreniere, Rene

Article Type: Research Article

Abstract: Employing passive immunization - using a heterologous anti-CD38 IgG antibody containing serum - in SCID mice injected subcutaneously with human multiple myeloma cells, we have shown that treatments with the antiserum - especially in the presence of complement - significantly decreased cancer growth. However, administered antibody and complement was not sufficient in amount to prevent cancer cell multiplication and cancer growth expansion to a satisfactory degree. Larger volumes of the same components more than likely would have further reduced cancer growth and prolonged the life of mice. In control mice, cancer growth progressed faster proving that lytic immune response against …multiple myeloma cells is necessary for cancer cell kill. Show more

Keywords: Cancer, modified vaccination technique, prophylactic, therapeutic

DOI: 10.3233/HAB-160295

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 53-57, 2016

Authors: Hamedani, Shima Yazdandoost | Taheri, Mohammad | Sajjadi, Elham | Omrani, Mir Davood | Mazdeh, Mehrdokht | Arsang-Jang, Shahram | Panah, Akram Sadat Tabatabaei | Sayad, Arezou

Article Type: Research Article

Abstract: BACKGROUND: Multiple sclerosis (MS) is an inflammatory, immune-mediated, demyelinating disease of the central nervous system which disturbs cerebral vascular permeability. OBJECTIVE: Matrix metalloproteinase can increase capillary permeability and are involved in neurological diseases. METHODS: The study compared the expression level of MMP9 gene in RRMS samples with normal individuals in Iran. RNA from the buffy coat of 50 RRMS patients and 50 normal controls were extracted. All patients were HLA-DRB1*15 negative and were responders to interferon-beta with a normal vitamin D level. The level of MMP9 gene expression was measured using …quantitative RT-PCR. RESULTS: The RRMS patients manifested a higher expression level of MMP9 than their normal counterparts (P= 0.02). Age-wise, there was no correlation between different age groups (> 30, 30-40, 40 <). In terms of sex, only the female patients manifested a statistically significant increase in MMP9 (p value = 0.037). Besides, there was no linear correlation between MMP9 expression level and risk of Expanded Disability Status Scale of Kurtzke (EDSS); nor were there any significant correlation between expression status of MMP9 and duration of the disease. CONCLUSION: Up regulation of MMP9 gene expression would happen in RRMS IFN-B responders, but the level of increase in female patients is significantly more than males. Show more

Keywords: MMP9, multiple sclerosis, expression, IFN-B

DOI: 10.3233/HAB-160292

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 59-64, 2016

Authors: Nazdik, Marzieh Khoshbin | Taheri, Mohammad | Sajjadi, Elham | Arsang-Jang, Shahram | Koohpar, Zeinab Kazaei | Inoko, Hidetoshi | Sayad, Arezou

Article Type: Research Article

Abstract: OBJECTIVES: Multiple sclerosis (MS) is an autoimmune disease involving the central nervous system (CNS) with unknown immunopathogenic mechanisms. Matrix metalloproteinase-9 (MMP-9) facilitates T-cell migration into the CNS while the tissue inhibitor matrix metalloproteinase-1 (TIMP-1) inhibits MMP-9 actions. The aim of this study was to evaluate the expression of TIMP-1 RNA and MMP-9/TIMP-1 RNA ratio in blood cells of Iranian patients with relapsing-remitting multiple sclerosis (RRMS) treated with IFNb. MATERIAL AND METHODS: The study compared the expression level of TIMP-1 gene in RRMS samples with normal individuals in Iran and the results were compared using a ratio …of MMP-9 to TIMP-1. All patients were HLA-DRB1*15 negative and were responders to interferon-beta with a normal vitamin D level. RESULTS: The RRMS patients manifested a lower expression level of TIMP-1 RNA than their normal counterparts although the result was not significant (P= 0.06). Also, the ratio of MMP-9 to TIMP-1 RNA increased significantly (P= 0.009). There was no linear correlation between TIMP-1 expression level and risk of Expanded Disability Status Scale of Kurtzke (EDSS); nor was there any significant correlation between expression status of TIMP-1 and duration of the disease. Although there was no significant decrease in TIMP-1 expression level, the MMP-9/TIMP-1 RNA ratio in RRMS was significantly higher than normal subjects. CONCLUSION: Further studies are recommended to compare MMP-9/TIMP-1 RNA ratio in patients before and after taking IFN-beta in order to find out if MMP-9/TIMP-1 RNA ratio can function as a proper marker of the bio efficacy of IFN-beta treatment of MS. Show more

Keywords: MMP-9, TIMP-1, expression, multiple sclerosis, real time PCR

DOI: 10.3233/HAB-160296

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 65-70, 2016

Authors: Rezaie, Zahra | Taheri, Mohammad | Kohan, Leila | Sayad, Arezou

Article Type: Research Article

Abstract: BACKGROUND: Multiple sclerosis (MS) as a complex neurological disease can be due to vitamin D deficiency. CYP27B1 is referred to as a vitamin D metabolizing enzyme. MATERIALS AND METHODS: This study compared the expression level of CYP27B1 in Relapsing-Remitting MS (RRMS) patients with normal individuals in Iran. The RNA was extracted from 50 RRMS patients and 50 normal controls. Quantitative RT-PCR was adopted to measure the expression level of CYP27B1 gene. RESULTS: The expression level of CYP27B1gene was significantly lower in the RRMS patients than their normal counterparts (P value = …0.04). Also, the RRMS females participating had a significant reduction in CYP27B1 gene expression compared to normal females (P-Value = 0.01). In addition, the correlation between CYP27B1 expression level, and the risk of Expanded Disability Status Scale of Kurtzke (EDSS) was not linear. Additionally, there was no significant correlation between expression status of CYP27B1gene and duration of the disease. CONCLUSION: A significant decrease in the expression level of CYP27A1 in female patients could indicate their greater vulnerability to MS than the male patients. Show more

Keywords: CYP27B1, expression, multiple sclerosis, real time PCR

DOI: 10.3233/HAB-160297

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 71-76, 2016

Authors: Esmati, Laya | Mehrabadi, Jalil Fallah | Bazaz, Masoumeh | Nejad, Hamideh Rouhani

Article Type: Research Article

Abstract: BACKGROUND: Human monoclonal antibodies are important molecules in clinical research. Current Limitations of mAb technologies namely instability of immortalized B-cell line and probability of forming unusual VH-VL pairs in phage-display method led to mAbs technology based on single plasma cell called ``SYMPLEX''. OBJECTIVE: In this method, cognate VH and VL fragments generated from individual antibody genes exactly the same as natural ones. METHODS: PBMCs of whole blood of an immunized candidate was used as a resource of rearranged Ab genes. Then flow-cytometric screening was performed to isolate VH and VL from PBMCs. Various …VH and VLκ were amplified by six pairs of primers. Overlap Extension PCR was accomplished to link VH and Vκ regions. ScFv inserted into T-vector and its sequence was determined and eventually analyzed by using blast analysis tools. RESULTS: Electrophoresis results indicated that VH and VL fragments were separately amplified by PCR with a length of about 400bp and linked through OE-PCR. Hence, ScFv, which was approximately 800bp in size, was constructed then sequencing and BLASTn results of the ScFv fragment consequently proved the accuracy. CONCLUSION: Results showed 88% similarity to available sequences in mentioned databank. ScFv was ultimately inserted into expression vector for producing recombinant human anti-tetanus mAb. Show more

Keywords: Overlap-extension PCR, ScFv, SYMPLEX technology

DOI: 10.3233/HAB-160298

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 77-83, 2016

Authors: Eftekharian, Mohammad Mahdi | Sayad, Arezou | Omrani, Mir Davood | Ghannad, Masoud Sabouri | Noroozi, Rezvan | Mazdeh, Mehrdokht | Mirfakhraie, Reza | Movafagh, Abolfazl | Roshanaei, Ghodratollah | Azimi, Tahereh | Inoko, Hidetoshi | Taheri, Mohammad

Article Type: Research Article

Abstract: BACKGROUND: Although Multiple Sclerosis (MS) is an autoimmune multifactorial disease with unknown etiology, various genetic and environmental factors are known to contribute to the pathogenesis of the disease. OBJECTIVE: Recent studies have confirmed that the suppressive function of regulatory T cells (T (reg)) is impaired in MS patients and that the FOXP3 gene is a crucial transcription factor in the regulation of CD4+ CD25+ FOXP3+ Treg cells. Polymorphisms in the promoter region of the FOXP3 gene may alter the gene expression level and, therefore, contribute to the disease susceptibility. …METHODS: The present study aimed to investigate the possible association between single nucleotide polymorphisms (SNPs) rs3761548 and rs2232365 in the FOXP3 gene and predisposition to MS. We conducted a case-control study on 410 patients with sporadic MS and 446 healthy controls. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Significant differences in distribution of both rs3761548 and rs2232365 A allele were found in MS patients in comparison to controls. Haplotype frequencies were also different among the studied groups. The A-A and C-G haplotype blocks showed a significant difference between case and controls. CONCLUSION: we have provided further evidence for the association between genetic variations and haplotypes in FOXP3 and MS in Iranian population. Show more

Keywords: Multiple sclerosis, FOXP3, single nucleotide polymorphism, Iranian population

DOI: 10.3233/HAB-160299

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 85-90, 2016

Article Type: Other

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 91-92, 2016

Article Type: Other

Citation: Human Antibodies, vol. 24, no. 3-4, pp. 93-, 2016