Mohamed I. Husseiny El-Sayed, Ph.D.

Mohamed Elsayed, Ph.D., started his academic career in 1994 in the Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Egypt. During that time he studied the susceptibility of bacteria and fungi to antibacterial and antifungal agents.

Soon after, he began his Ph.D. project at the Institute for Clinical Microbiology, Immunology and Hygiene, Friedrich-Alexander University, Erlangen-Nürnberg, Germany, in the laboratory of Dr. Michael Hensel. The ultimate goal of his Ph.D. project was the development of novel vaccination strategies using attenuated Salmonella as live carriers for recombinant vaccines. The idea was to utilize Salmonella to create recombinant live vaccines for mucosal immunization. This work provides a promising platform for the production of diverse vaccines

After obtaining his PhD, he went back to Egypt to start his faculty position as Assistant Professor of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Egypt.

Later he suspended his work in Egypt and started his work in the USA as a postdoctoral fellow at LA-Biomed at Harbor-UCLA Medical Center, Division of Infectious Diseases, Department of Medicine, Torrance, CA. He is involved in the project of generating isogenic null mutants of rFTR1 in R. oryzae to study the role of iron-related virulence, and also the expression and secretion of rFTR1 protein by yeast for vaccination purposes.

Dr. Elsayed then became a postdoctoral fellow in the laboratory of Dr Robert Seeger at Children Hospital Los Angeles, Division of Hematology/Oncology, Keck School of Medicine, USC, Los Angeles, CA. The goal of his project was to develop a novel vaccine against cancer cells using SPI2-T3SS of Salmonella as a live vector vaccine. We have constructed a new SPI2-based oral Salmonella vaccine against Survivin (TAA) and evaluated the vaccine immunogenicity and anti-tumor efficacy in two murine models of cancer. The combined use of Salmonella-based Survivin with GSL1 leads to enhanced immunogenicity and increased anti-tumor efficacy. This suggests that NKT ligands could be used as adjuvants for Salmonella-based cancer vaccines. Also, he examined the potential use of SPI2-T3SS of Salmonella as an oral vaccine against neuroblastoma. The rationale was based on our preliminary data that demonstrated a specific CD8 immune response after oral vaccination with Salmonella.

He then became a postdoctoral fellow in City of Hope with Dr. Simon Lacey, Division of Translational Vaccine Research, Beckman Research Institute. His project focused on the development of the concept of engineering recombinant BK virus (rBKV) to express HIV antigenic polypeptides as a candidate HIV vaccine. He used a novel strategy to generate a panel of recombinant BK virus (rBKV) expressing various fragments of the HIV-1 gag polypeptide and green fluorescent protein (GFP) under the control of the BKV late promoter. He has established a method for the generation of infectious rBKV from genomic DNAs introduced into cultured cells.

Currently, he is working as an Assistant Research Professor with Dr. Kevin Ferreri, Department of Diabetes and Metabolic Diseases Research, Beckman Research Institute at City of Hope. His project focuses on the development of an assay for the detection of circulating beta cell death in early onset Type 1 Diabetes using methylation-specific PCR. The onset of metabolic dysregulation in Type 1 Diabetes occurs after the autoimmune destruction of 60% to 80% of pancreatic insulin-producing beta cells. In addition, he implemented another novel idea for the production of a vaccine against Type 1 Diabetes using Salmonella as a live vector vaccine.

Development of oral Salmonella-based antigen-specific immunotherapy for T1DM:

The first project is the development of vaccine for type 1 diabetes. This project was initiated based on my PhD training work concerning the use of attenuated Salmonella typhimurium as an oral carrier of specific antigens. The data showed, oral vaccination prevented disease in the majority of animals and was accompanied by stabilization of blood glucose levels and induction of a tolerogenic cytokine profile. This is the first demonstration that Salmonella can be used as a tolerogenic antigen-specific vaccine, where an autoantigen of choice is expressed under the control of SPI2 in a regulated fashion inside the host antigen-presenting cells and that this can be combined with Salmonella-mediated expression of specific cytokines by host cells for induction of tolerance. These findings were published in peer reviewed journal (Vaccine 2014, 32(20), 2300-2307).

The aims of the study:

1- Investigate the mechanism of autoantigen-specific therapy on the immune system.

2- Determine the effect of multiple autoantigens on prevention of T1D.

3- Determine the effect of an alternative immunomodulatory on prevention of T1D.

4- Investigate the effect of anti-CD3 combination therapy on the immune system of T1D

5- Explore the effect of oral Salmonella-based vaccine on reversal of established T1D.

Methylation-specific PCR for diabetic biomarkers:

The second project is the development of a methylation-specific PCR assay for quantifying levels of unmethylated mouse and human insulin genes. This study is primarily aimed at development of a method for early diagnosis of beta cell death at the onset of type 1 diabetes. The development of the assay for the mouse model completed and published this data in peer reviewed journal (PLOS ONE, 2012, 7(10), e47942) describing the assay and its ability to detect beta cell death before the rise in blood glucose levels. In addition the development of the human assay is already done, and the application to clinical samples obtained from islets transplant patients and the finding results published in (PLOS ONE 2014, 9(4), e94591). The current focus is validation of the assay by monitoring beta cell loss during the early “honeymoon” phase of T1D in humans. The natural history of disease progression in patients monitored by collecting longitudinal samples throughout the first year post-diagnosis, and comparison with metabolic data and autoantibody titers and the finding results will publish soon (as Manuscript under preparation).

Validation of the assay will allow investigation of even earlier time points, such as within the high disease risk population. Additionally, the assay will be applicable to effectively monitor beta cell death for assessment of pre-diabetic treatment regimens. Currently we developed qMSP assay using two-steps for PCR and starting Bi-PAP technology to optimize the sensitivity of the assay.

Pyrophosphorolysis-Activated Polymerization (PAP) technology for monitoring of islet graft after transplantation:

The third project is the monitoring of the islet graft through detection of donor specific DNA markers in recipient plasma using Pyrophosphorolysis-Activated Polymerization (PAP) technology. The goal of this study is to develop a sensitive and specific assay for monitoring islet graft injury based on the hypothesis that damaged donor islet cells release their DNA into recipient circulation. Therefore the presence of donor-specific DNA or its increased levels in recipient’s plasma would indicate islet graft injury. In the assay, eleven common single nucleotide polymorphisms (SNPs) were selected to differentiate donor from recipient DNA with a theoretical power of 99.4%. 22 PAP based assays were developed to detect the two alleles of each selected SNP. In the current study, a robust method with a high sensitivity and specificity were developed for the accurate detection and quantitation of the low copy number of circulating donor DNA in islet transplant recipient’s plasma.

Patents:

Husseiny MI and Ferreri K. Compositions and methods for detecting unmethylated DNA (TEC 13-043); ML Ref. No.: 48440-566P01US. (Provisional application), 2015.

Husseiny MI and Ferreri K. Attenuated salmonella bacteria and methods of using (Patent: WO2014/144965, PCT/US2014/029591), 2014.