Enhancing soluble expression and purification of a bispecific scFv antibody in Escherichia coli using chaperones Zahra Alidousti-Shahraki, Afrooz Amini-Dehkordi, Nafiseh Esmaeil, Vajihe Akbari Research in Pharmaceutical Sciences, 2026 Background and purpose: The use of a bispecific antibody engineered as a tandem single-chain fragment variable antibody (scFv-BsAb) offers several advantages over full-length antibodies, including enhanced tissue penetration and the potential for production in bacterial cells. However, production of them in bacteria usually results in the formation of inclusion bodies. The current work aimed to facilitate proper folding and soluble expression of a recombinant scFv-BsAb targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) by co-expression with molecular chaperones. Experimental approach: Escherichia coli BL21(DE3) cells harboring pET28a-scFv-BsAb and pKJE7 plasmids were used as host cells for protein expression. The effect of different temperatures and IPTG concentrations on the expression of soluble protein with or without expression of chaperones was investigated by SDS-PAGE and densitometry analysis. Subsequently, the expressed protein was subjected to native purification using a Ni-NTA affinity column. Then, the binding ability of the scFv-BsAb to the receptors on peripheral blood mononuclear cells (PBMCs) was evaluated by flow cytometry. Findings/Results: Co-expression with chaperones slightly enhanced the expression of soluble scFv-BsAb. The optimal condition for soluble expression of protein is at a temperature of 30 °C and an IPTG concentration of 1 mM when co-expressed with chaperones. The final yield of protein in co-expression with chaperones was 2.9 mg protein per 1 L of culture. Conclusion and implications: Optimization of culture conditions and co-expression with molecular chaperones may be useful for the production of soluble and functionally bioactive scFv antibodies in E. coli.
Liposomal AZD5363 Displays Antiproliferation Activities and Induces Apoptosis on Y79 Retinoblastoma Cancer Cells Zahra Khabazian, Nafiseh Esmaeil, Maryam Khanehzad, Amir Hossein Naderi Majd, Mahdi Tohidian, et al. South Asian Journal of Cancer, 2025 Objectives Retinoblastoma (RB) is an aggressive intraocular cancer that usually develops during infancy and childhood. As an Akt kinase inhibitor, AZD5363 is a novel drug whose encapsulation into liposomes enhances its bioavailability and biomedical potential. In the present study, a liposomal membrane was created around AZD5363 to assess its efficacy on the Y79 cancer cell line. Materials and Methods AZD5363 nanoparticles were synthesized by the thin film hydration method. Dynamic light scattering (DLS) and field emission scanning electron microscopy (FESEM) techniques were applied to evaluate the particle size, and the morphology of the liposomal AZD5363 (Lipo-AZD5363). The MTT test was used to assess the half maximal inhibitory concentration (IC50) of Lipo-AZD5363, and the cytotoxic effects of Lipo-AZD5363 and doxorubicin (Dox) were investigated on the Y79 cell line. Flow cytometry was used to study apoptotic induction in selected groups. Also, the PTEN/AKT/FOXO1 gene expression level was measured using quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. Results Treatment with Lipo-AZD5363 inhibited the proliferation of Y79 RB cancer cell line in a dose-dependent manner. Lipo-AZD5363, at a lower concentration, was significantly more cytotoxic than Dox in terms of enhanced cell death (p < 0.05).Furthermore, flow cytometry showed that Lipo-AZD5363 and Dox induce apoptosis in these cells. However, the number of apoptotic cells in the Lipo-AZD5363 group was clearly higher than that in the Dox group (p < 0.001). Real-time PCR analysis indicated that Lipo-AZD5363 treatment resulted in an increase in PTEN and FOXO1 gene expression and a decrease in AKT gene expression. Our study revealed that all results were statistically more significant in the Lipo-AZD5363 group than in the Dox group (p < 0.01, <0.01, and <0.001, respectively). Conclusion Lipo-AZD5363 inhibits proliferation and promotes apoptosis of RB cells by inhibiting the PI3K/AKT signaling pathway. Thus, Lipo-AZD5363 may be a promising candidate for cancer therapy. However, more experimental evidence is needed for its use in the pharmacological treatment of RB.
Emerging Mechanisms of NK Cell Dysfunction in Endometriosis: The Role of Autophagy, Metabolism, Cytokines, Exosomes, and Trogocytosis Mohammad Abbaszadeh, Mojdeh Soltani, Sara Falahi, Nafiseh Esmaeil American Journal of Reproductive Immunology, 2025 Endometriosis (EMS) is a persistent, inflammatory condition that relies on estrogen and is distinguished by the proliferation of endometrial tissue outside the confines of the uterus. The impact on the well‐being of individuals affected can be significant, as it is linked to pelvic pain and reduced fertility in women of reproductive age. Over 30 years have passed since initially discovering a malfunction in the activity of natural killer (NK) cells in individuals with EMS. Several aspects that contribute to NK cell dysfunction have been explored by researchers over the years, such as the upregulation of inhibitory receptors, downregulation of activating receptors, and the exhaustion process. Nonetheless, there are still many aspects that have yet to be identified. The objective of this review is to explore whether there is a connection between mechanisms that have not yet been explored and the malfunctioning of EMS‐derived NK cells. Autophagy, metabolism, trogocytosis, tunneling nanotubes (TNT), and exosomes are among the factors that play a role in these processes. The primary objective of this publication is to provide valuable insights for future research on NK cells, with the aim of enhancing our understanding of the disease's causes and identifying more effective targets for immunotherapy.
Different behavior of NK cells isolated from healthy women and women with recurrent spontaneous abortion after treatment with human amniotic epithelial cells Fatemeh Rezayat, Nafiseh Esmaeil, Parvaneh Nikpour, Awat Feizi, Abbas Rezaei Journal of Leukocyte Biology, 2025 Maternal immunotolerance during pregnancy is heavily dependent on the critical properties of human amniotic epithelial cells (hAECs). Recurrent spontaneous abortion (RSA) is one of the most common diseases in women and is caused by feto-maternal immunotolerance disruption. The objective of this study is to investigate how hAEECs affect pNK cells isolated from RSA and healthy women in terms of immunomodulation. Peripheral blood NK cells were isolated from 20 women with RSA and 20 healthy women. Purified NK cells were co-cultured with hAECs, obtained from full-term healthy pregnant women at different cellular ratios. After 24 and 72 h of incubation, the expression of immunomodulatory genes in hAECs, immunophenotype, and cytotoxicity of NK cells, and cytokine production were investigated using real-time PCR, flow cytometry, and ELISA techniques, respectively. We observed a significant increase in TGF-β and IL-10 production, and CD56bright CD16+ subpopulation in pNK cells, a significant decrease in IFN-γ production and CD107a and FasL expression on NK cells. Also, NK cells' cytotoxicity against K562 cells was diminished after co-culture with hAECs. The expression of TGF-β and HLA-G genes by hAECs was diminished after co-culture with NK cells isolated from women with RSA. Our research indicates that the interaction between NK cells and hAECs influences the phenotype and function of both cells. Also, NK cells belonging to women with RSA and healthy women exhibit different behavior during treatment with hAECs, possibly due to NK cell dysfunction. However, extensive research is required to assess NK cell defects and their mutual interaction with hAECs.
Optimization of in vitro expansion and activation of human natural killer cells against breast cancer cell line Avicenna Journal of Medical Biotechnology, 2020
Decreased serum levels of interleukin-35 among multiple sclerosis patients may be related to disease progression Journal of Biological Regulators and Homeostatic Agents, 2018
Cytokine indexes in pemphigus vulgaris: Perception of its immunpathogenesis and hopes for non-steroidal treatment Iranian Journal of Pharmaceutical Research, 2017
Comparison of B-cells differentiation to plasmablasts at presence of anti-human CD40 and anti-immunoglobulin M f (Ab)'2 or lipopolysaccharide and anti-human CD40 stimulators (in-vitro) Journal of Isfahan Medical School, 2017
Assessment of the expression level of B-cell lymphoma 6 (BCL6) in peripheral blood CD38+ B lymphocytes in patients with common variable immunodeficiency (CVID) Journal of Isfahan Medical School, 2017
Evaluation of soluble human leukocyte antigen-G (sHLA-G) isoforms and regulatory T cells in relapsing-remitting multiple sclerosis Iranian Journal of Allergy Asthma and Immunology, 2015
Study of carbon nano-tubes effects on the chondrogenesis of human adipose derived stem cells in alginate scaffold International Journal of Preventive Medicine, 2014
Suppressive impact of anethum graveolens consumption on biochemical risk factors of atherosclerosis in hypercholesterolemic rabbits International Journal of Preventive Medicine, 2013
Comparison between chondrogenic markers of differentiated chondrocytes from adipose derived stem cells and articular chondrocytes in vitro Iranian Journal of Basic Medical Sciences, 2013
The apoptotic effects of sirtuin1 inhibitor on the MCF-7 and MRC-5 cell lines Research in Pharmaceutical Sciences, 2013
Comparison of phenotypic characterization between differentiated osteoblasts from stem cells and calvaria osteoblasts in vitro International Journal of Preventive Medicine, 2013
Comparison of the production of extracellular matrix in nucleus pulposus of intervertebral disc in alginate and chitosan-gelatin scaffolds Journal of Isfahan Medical School, 2013
The role of maternal thyroid stimulating hormone receptor blocking antibodies in the etiology of congenital hypothyroidism in Isfahan, Iran International Journal of Preventive Medicine, 2012
Serum monocyte chemotactic protein-1 and nitric oxide in cutaneous leishmaniasis patients before and after treatment with glucantime Journal of Isfahan Medical School, 2012
Serum monocyte chemotactic protein-1 and nitric oxide in cutaneous leishmaniasis patients before and after treatment with glucantime Journal of Isfahan Medical School, 2012
IFN-γ, IL-4 and IL-17 concentration in tumor tissue, adjacent tumor tissue and the serums of patients with breast cancer Journal of Isfahan Medical School, 2011
The role of maternal tsh receptor blocking antibody in the etiology of congenital hypothyroidism in isfahan Journal of Isfahan Medical School, 2011
Phenotypic characterizations and comparison of adult dental stem cells with adipose-derived stem cells International Journal of Preventive Medicine, 2010
Immune Checkpoint Blockade and Emerging Combination Platforms in Breast Cancer: A Narrative Review M Abbaspour, N Esmaeil, V Akbari, M Ghoreishi, H Rafiepoor Breast Cancer: Targets and Therapy, 588323 , 2026 2026
Enhancing soluble expression and purification of a bispecific scFv antibody in Escherichia coli using chaperones Z Alidousti-Shahraki, A Amini-Dehkordi, N Esmaeil, V Akbari Research in Pharmaceutical Sciences 21 (3), 323-332 , 2026 2026
Rethinking MS Therapeutics: From Disease Pathogenesis Mechanisms to AI-Driven Drug Discovery M Ziaei, M Sehhati, M Torabi, N Esmaeil, F Ghasemi Journal of Neuroimmune Pharmacology 21 (1), 21 , 2026 2026
High intensity functional training versus traditional resistance training effects on inflammatory, metabolic, and physical outcomes in overweight men a randomized controlled trial F Hosseini Moshkenani, S Abedi, F Shabkhiz, R Soori, N Esmaeil Scientific Reports , 2026 2026
MicroRNA-mediated post-transcriptional regulation in the pathogenesis of multiple sclerosis M Ziaei, L Dayani, N Esmaeil, HT Ghashghaei, M Aliomrani Neuroscience and Behavioral Physiology, 1-35 , 2026 2026
Tregs in Allogeneic Hematopoietic Stem Cell Transplantation: Suppressing Graft-Versus-Host Disease while Preserving Graft-Versus-Leukemia NK Nekouei, N Najjari, H Farajifard, N Esmaeil, M Naseroleslami, S Sari, ... Transplantation and Cellular Therapy , 2025 2025
Fast Fitness: comparing a 7-minute high-intensity interval training vs. walking for cardio-metabolic health in overweight women V Hovsepian, N Soltani, SM Marandi, M Yazdi, N Esmaeil 2025
Emerging Mechanisms of NK Cell Dysfunction in Endometriosis: The Role of Autophagy, Metabolism, Cytokines, Exosomes, and Trogocytosis M Abbaszadeh, M Soltani, S Falahi, N Esmaeil American Journal of Reproductive Immunology 94 (2), e70138 , 2025 2025
Different behavior of NK cells isolated from healthy women and women with recurrent spontaneous abortion after treatment with human amniotic epithelial cells F Rezayat, N Esmaeil, P Nikpour, A Feizi, A Rezaei Journal of Leukocyte Biology 117 (4), qiaf020 , 2025 2025 Citations: 3
Galectin-9 induces IL-1β production as a key inflammatory cytokine in the acute myeloid leukemia cell line (U937) N Behranvand, F Nasri, MV Moghaddam, MD Gholami, E Safari, ... Research in pharmaceutical sciences 20 (2), 304-315 , 2025 2025 Citations: 2
DCs pulsed with hypochlorous acid-treated tumor cell lysates present antigens efficiently and induce CD8+ T cell activation through cross-presentation M Abbaspour, N Esmaeil, M Rezazadeh, M Minaiyan, V Akbari Research in pharmaceutical sciences 20 (2), 230-240 , 2025 2025 Citations: 2
Synthesis of a covalently linked bismuthene–graphene heterostructure loaded with mitomycin C for combined radio-thermo-chemotherapy of triple-negative breast cancer M Varzandeh, M Karbasi, S Labbaf, J Varshosaz, N Esmaeil, R Birang Journal of Materials Chemistry B 13 (26), 7769-7784 , 2025 2025 Citations: 4
The niosomal nelarabine as a promising nano combination for retinoblastoma treatment: an in vitro study—experimental research M Tohidian, N Esmaeil, M Soleimani, M Khanehzad, AHN Majd, ... Annals of Medicine and Surgery 87 (1), 121-129 , 2025 2025
Liposomal AZD5363 displays antiproliferation activities and induces apoptosis on Y79 retinoblastoma cancer cells Z Khabazian, N Esmaeil, M Khanehzad, AHN Majd, M Tohidian, ... South Asian Journal of Cancer , 2024 2024 Citations: 1
Evaluation of the natural killer cell subsets and their relationship with serum interferon gamma and vitamin D levels in women with stages III and IV endometriosis: A case … SN Chamgordani, N Esmaeil, M Hashemi, A Amari, M Seyedtabib, ... International Journal of Reproductive Biomedicine 22 (7), 593 , 2024 2024 Citations: 2
More balance toward activating receptors and cytotoxic activity of NK cells ex vivo differentiated from human umbilical cord blood-derived CD34+ stem cells in comparison with … F Ghaedrahmati, N Esmaeil, V Akbari, F Ashrafi Heliyon 10 (15) , 2024 2024 Citations: 6
Strong capacity of differentiated PD-L1 CAR-modified UCB-CD34 + cells and PD-L1 CAR-modified UCB-CD34 + -derived NK cells in killing target cells and … F Ghaedrahmati, V Akbari, H Seyedhosseini-Ghaheh, N Esmaeil Stem Cell Research & Therapy 15 (1), 257 , 2024 2024 Citations: 7
Decrease of Tregs cells and increase of exhausted Treg cells as the predictors of COVID19 severity SM Mirniam, A Andalib, M Radandish, R Sami, N Esmaeil Journal of Clinical Virology Plus 4 (3), 100189 , 2024 2024 Citations: 3
Investigating the effect of Fusobacterium nucleatum on the aggressive behavior of cancer-associated fibroblasts in colorectal cancer F Eskandari-Malayeri, M Rezeai, T Narimani, N Esmaeil, M Azizi Discover Oncology 15 (1), 292 , 2024 2024 Citations: 11
Pathway-based Mendelian randomization for pre-infection IL-6 levels highlights its role in coronavirus disease Z Kamali, N Esmaeil, CHL Thio, A Vaez, H Snieder Genes 15 (7), 889 , 2024 2024 Citations: 1
MOST CITED SCHOLAR PUBLICATIONS
Roles of GM-CSF in the pathogenesis of autoimmune diseases: an update N Lotfi, R Thome, N Rezaei, GX Zhang, A Rezaei, A Rostami, N Esmaeil Frontiers in immunology 10, 1265 , 2019 2019 Citations: 278
Silymarin impacts on immune system as an immunomodulator: One key for many locks N Esmaeil, SB Anaraki, M Gharagozloo, B Moayedi International immunopharmacology 50, 194-201 , 2017 2017 Citations: 185
Suppressive impact of anethum graveolens consumption on biochemical risk factors of atherosclerosis in hypercholesterolemic rabbits M Setorki, M Rafieian-Kopaei, A Merikhi, E Heidarian, N Shahinfard, ... International Journal of Preventive Medicine 4 (8), 889 , 2013 2013 Citations: 112
Dust events, pulmonary diseases and immune system N Esmaeil, M Gharagozloo, A Rezaei, G Grunig American journal of clinical and experimental immunology 3 (1), 20 , 2014 2014 Citations: 111
Perspective: ambient air pollution: inflammatory response and effects on the lung's vasculature G Grunig, LM Marsh, N Esmaeil, K Jackson, T Gordon, J Reibman, ... Pulmonary circulation 4 (1), 25-35 , 2014 2014 Citations: 109
A randomized double‐blind, placebo‐controlled study of therapeutic effects of silymarin in β‐thalassemia major patients receiving desferrioxamine B Moayedi, M Gharagozloo, N Esmaeil, MR Maracy, H Hoorfar, ... European journal of haematology 90 (3), 202-209 , 2013 2013 Citations: 83
Phenotypic characterizations and comparison of adult dental stem cells with adipose-derived stem cells R Alipour, F Sadeghi, B Hashemi-Beni, SH Zarkesh-Esfahani, F Heydari, ... International journal of preventive medicine 1 (3), 164 , 2010 2010 Citations: 73
Effects of apple juice on risk factors of lipid profile, inflammation and coagulation, endothelial markers and atherosclerotic lesions in high cholesterolemic rabbits M Setorki, S Asgary, A Eidi, AH Rohani, N Esmaeil Lipids in health and disease 8 (1), 39 , 2009 2009 Citations: 72
Evaluation of the in vivo acute antiinflammatory response of curcumin-loaded nanoparticles M de Almeida, BA da Rocha, CRL Francisco, CG Miranda, ... Food & function 9 (1), 440-449 , 2018 2018 Citations: 63
Method and key points for isolation of human amniotic epithelial cells with high yield, viability and purity H Motedayyen, N Esmaeil, N Tajik, F Khadem, S Ghotloo, B Khani, ... BMC research notes 10 (1), 552 , 2017 2017 Citations: 62
Immunosuppressive effect of silymarin on mitogen‐activated protein kinase signalling pathway: The impact on T cell proliferation and cytokine production M Gharagozloo, S Jafari, N Esmaeil, EN Javid, B Bagherpour, A Rezaei Basic & clinical pharmacology & toxicology 113 (3), 209-214 , 2013 2013 Citations: 59
Evaluation of the effect of GM-CSF blocking on the phenotype and function of human monocytes N Lotfi, GX Zhang, N Esmaeil, A Rostami Scientific reports 10 (1), 1567 , 2020 2020 Citations: 58
Oxidative stress and age-related changes in T cells: is thalassemia a model of accelerated immune system aging? M Ghatreh-Samani, N Esmaeili, M Soleimani, M Asadi-Samani, ... Central European Journal of Immunology 41 (1), 116-124 , 2016 2016 Citations: 58
An 8-year study of people with multiple sclerosis in Isfahan, Iran: association between environmental air pollutants and severity of disease F Ashtari, N Esmaeil, M Mansourian, P Poursafa, O Mirmosayyeb, ... Journal of neuroimmunology 319, 106-111 , 2018 2018 Citations: 56
Targeting immune checkpoints: how to use natural killer cells for fighting against solid tumors F Ghaedrahmati, N Esmaeil, M Abbaspour Cancer Communications 43 (2), 177-213 , 2023 2023 Citations: 53
Serum levels of TGFβ, IL-10, IL-17, and IL-23 cytokines in β-thalassemia major patients: the impact of silymarin therapy S Balouchi, M Gharagozloo, N Esmaeil, M Mirmoghtadaei, B Moayedi Immunopharmacology and immunotoxicology 36 (4), 271-274 , 2014 2014 Citations: 50
Comparison between chondrogenic markers of differentiated chondrocytes from adipose derived stem cells and articular chondrocytes in vitro M Mardani, B Hashemibeni, MM Ansar, SHZ Esfahani, M Kazemi, ... Iranian journal of basic medical sciences 16 (6), 763 , 2013 2013 Citations: 50
The role of distinct subsets of macrophages in the pathogenesis of MS and the impact of different therapeutic agents on these populations M Radandish, P Khalilian, N Esmaeil Frontiers in immunology 12, 667705 , 2021 2021 Citations: 49
Interleukin 13–and interleukin 17A–induced pulmonary hypertension phenotype due to inhalation of antigen and fine particles from air pollution SH Park, WC Chen, N Esmaeil, B Lucas, LM Marsh, J Reibman, G Grunig Pulmonary circulation 4 (4), 654-668 , 2014 2014 Citations: 44
The exercise training modulatory effects on the obesity-induced immunometabolic dysfunctions N Soltani, SM Marandi, M Kazemi, N Esmaeil Diabetes, Metabolic Syndrome and Obesity, 785-810 , 2020 2020 Citations: 42
