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Ghoneim, N. H., K. A. Abdel-moein, A. M. BARAKAT, A. G. Hegazi, K. A. A. El-Razik, and S. A. SADEK, "Isolation and molecular characterization of Campylobacter jejuni from chicken and human stool samples in Egypt", food science and technology, vol. 41, issue 1, pp. 195-202, 2021.
El-Leithy, M. A., W. D. Saleh, and H. M. Fathy, Isolation and insecticidal activity of Bacillus thuringiensis strains obtained from North Sinai, Egypt., , 2004. Abstract
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El-Houseini, M. E., M. M. Sleem, A. M. Bahnassy, K. M. Shaaban, T. K. Saber, M. El-Sherbini, and T. D. Hussein, "Isolation and Immuno-Phenotypic Characterization of Tumor Infiltrating Lymphocytes (TILs) Obtained from Breast Malignant Tumor Tissues of Egyptian Patients.", Journal of the Egyptian National Cancer Institute, vol. 20, issue 4, pp. 379-86, 2008 Dec. Abstract

BACKGROUND: Isolation and phenotypic characterization of tumor infiltrating lymphocytes (TILs) in some malignant tumors have been shown. TILs possess a good prognostic value as well as a therapeutic effect in these solid tumors. Our preliminary work shed some light on a good possibility of synthesis and secretion of specific protease enzyme system with a dimeric structure above 92 kDa for the lytic activity of TILs against breast tumor cells propagated ex vivo.

PURPOSE: This work aims at first isolation, activation and immuno-phenotypic characterization of TILs derived from malignant tumor tissues of breast cancer patients. Second, to optimize the conditions for the biological therapeutic efficiency of the identified TILs subpopulations as targeted cell therapy against breast cancer.

PATIENTS AND METHODS: The present work presented twelve patients with breast cancer from NCI, Cairo. Tcell isolation, activation, immunophenotyping and immunohistochemical investigations were performed. Enzymatic digestion method, mesh with pore size 355 & 45mm and flow cytometric analysis were used.

RESULTS: The results revealed that, lymphocytes infiltrating the malignant tumor tissues were mainly of the Tcell type indicated by CD45RO positive markers as shown by immunohistochemical observations. The immunophenotypic analysis of the isolated TILs obtained from breast tumor tissues specimens and activated with interleukin- 2 (IL-2), showed that the ratio of CD4+/CD8+ was 0.89 which represents helper and cytotoxic sub-populations of TILs, respectively. Meanwhile, the ratio of CD4+/CD25+ was 16.03 representing the regulatory system of TILs subpopulation. In the peripheral blood of patients, the percentages of the CDs positive cells were different and the ratio of CD4+/CD8+ was 1.14+/-0.57 whereas the ratio of CD4+/CD25+ was 18.38+/-5.95. After mixing the isolated TILs and the T-lymphocytes obtained from the peripheral blood, the ratio of CD4+/CD8+ increased insignificantly to 1.45+/-0.67. Also the ratio of CD4+/CD25+ increased rough insignificantly, to 23.64+/-9.83.

CONCLUSION: The isolated and identified TILs subpopulations have to be tested for their biological therapeutic efficacy first at ex-vivo level using the cr51 release assay; second at in-vivo level using experimental animal models as a sub-clinical investigation before going further to clinical study of using TILs as targeted bio-immunotherapy against human cancers.

KEY WORDS: TILs - Immunohistochemistry - Immunophenotyping - Breast cancer.

Bashir, H. S., A. M. Mohammed, M. A.M., and S. A.S., "Isolation and Identification of Two Flavonoids from Acaca Nitotica (Leguminosae) Leaves", journal of forest products and industries, vol. 3, issue 5, pp. 211-215, 2014. paper_acacia_seham.pdf
Hamed, A. H., O. - H. M. El-Banna, G. A. M. Ghanem, H. Elnagaar, and M. S. Shafie, "Isolation and Identification of Tobacco rattle tobravirus Affecting Onion (Allium cepa L.) Plants in Egypt.", International Journal of Virology, vol. 8, issue 1, 2012. Abstract
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Hamed, A. H., M. Om-Hashim, El-Banna, G. A. M. Ghanem, H. Elnagaar, and M. S. Shafie, "Isolation and identification of Tobacco rattle Tobravirus affecting onion (Allium cepa L.) plants in Egypt. ", International Journal of Virology, vol. 8 , issue 1, pp. 39-49, 2012.
Hamed, A. H., E. - B. O. - H. M., G. G. A.M., H. El-Nagaar, and M. S. Shafie, "Isolation and Identification of Tobacco Rattle Tobravirus affecting onion (Allium cepa L.) in Egypt.", International Journal of Virology, vol. 8, issue 1, pp. 39-49, 2012. Abstract
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Hamed, A. H., O. - H. M. El-Banna, G. A. M. Ghanem, H. Elnagaar, and M. S. Shafie, "Isolation and Identification of Tobacco Rattle Tobravirus affecting onion (Allium cepa L.) in Egypt. ", International Journal of Virology, vol. 8, issue 1, pp. 39-49, 2012. Abstract

Tobacco Rattle Virus (TRV) was isolated from naturally infected onion (Allium cepa L.) plants growing in the fields of onion plants during the survey carried out in two successive seasons (2007-2008 and 2008-2009) in seven Egyptian Governorates. Plants showing yellowing, malformation, yellow stripping, white necrotic stripes and stunting symptoms were collected and subjected to identification studies that based on host range, symptomatology, modes of transmission, serological tests, inclusion bodies and morphology of virus particles. The virus was transmitted by mechanical inoculation and by seeds. On the other hand, cytological changes accompanied with the infection were investigated. Different methods of serological detection of the virus were also tested. The obtained results indicated that the host range of the expanded to 7 different plant families. The virus was transmitted mechanically and by seeds with percentages ranged between 8-13%. Different serological methods were used successfully for detection of TRV i.e., DAS-ELISA, TBIA and DIBA. Infection of tobacco leaves with TRV resulted in the formation of amorphous inclusion bodies in the cytoplasm. Light microscopy examination of semi thin sections in both healthy and artificially infected onion leaves showed anatomical changes which reflecting on the external symptoms on infected plants. Also, electron microscopy observed tubular particles with two main dimensions (length 48-114 nm and 180-197 nm and 22 nm width). Investigation of ultrathin sections by transmission electron microscopy revealed changes in both nucleus and chloroplast. According to the available data, TRV was isolated and identified for the first time in Egypt from onion plants during the present study.

Hemaia, M., L. Motawe, F. M. Ibrahim, M. E. Ibrahim, E. A. MAHMOUD, and H. F. Aly, "Isolation and Identification of Terpenoids and Sterols of Nepeta cataria L.", International Journal of PharmTech Research, vol. 8, issue 10, pp. 10-17, 2015. Abstract
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Farouk, M. M., A. A. E. Molla, D. G. Abo El-Hassan, and A. I. Tanios, "Isolation and identification of salmonella spp. From diarrheic sheep and goats in Egypt", J. Egypt. Vet. Med. Ass., vol. 76, issue 4, pp. 485-496, 2016.
Kamil, Z., M. Saleh, and S. Moustafa, Isolation and Identification of Rhizosphere Soil Chitinolytic Bacteria and their Potential in Antifungal Biocontrol 1, : Citeseer, 2007. Abstractisolation_and_identification_of_rhizosphere_soil.pdf

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Khalil, E. M., M. F. Attia, E. A. Salama, A. A. Kishtah, and O. M. El-Banna, "Isolation and identification of potyvirus from naturally infected potato in Egypt", Egyptian Journal of Phytopathology (Egypt), 1986. Abstract
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Tharwat, N. A., "Isolation and identification of pectinolytic fungi isolated from soil amended with pectin", . Egypt. J. Bot.,, vol. 52, issue (1), pp. : 117-127., 2012.
Moussa, T. A. A., and D. M. I. Ali, "Isolation and Identification of Novel Dissaccaride of α- L- Rhamnose from Penicillium chrysogenum", World Applied Sciences Journal, vol. 3, issue 3, pp. 476-486, 2008.
Moussa, T. A. A., and D. M. I. Ali, "Isolation and Identification of Novel Disaccharide of $ąppa$-L-Rhamnose from Penicillium chrysogenum", World Applied Sciences Journal, vol. 3, no. 3, pp. 476–486, 2008. Abstract
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Castañeda Vázquez, H., S. Jäger, W. Wolter, M. Zschöck, C. Vazquez, and A. El-Sayed, Isolation and identification of main mastitis pathogens in Mexico, , vol. 65, issue 2: Escola de Veterinária UFMG, pp. 377 - 382, 2013. Abstract
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El-Assal, S. E. - D., N. A. Youssef, and G. A. Amin, "Isolation and identification of locally isolated bacterial strains effective against whitefly Bemisia tabaci", Archives of Agronomy and Soil Science, vol. 59, issue 6: Taylor & Francis, pp. 779-790, 2013. Abstract
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Youssef, N. A., G. A. Amin, and S. E. El-Assal, "Isolation and Identification of Local Isolated Bacterial Strains as a Biological Control Agent against the Cotton Leaf Worm, Spodoptera littoralis (Boisd.)", Egyptian Journal of Biological Pest Control, vol. 24, issue 1: Egyptian Society for Biological Control of Pests, pp. 229, 2014. Abstract
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Elzeini, H. M., A. -rhman A. -atti Ali, N. F. Nasr, Y. E. Elenany, and A. A. M. Hassan, "Isolation and identification of lactic acid bacteria from the intestinal tracts of honey bees, Apis mellifera L., in Egypt", Journal of Apicultural Research, vol. 60, issue 2, pp. 1-9, 2021.
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Elzeini, H. M., A. -rhman A. -atti Ali, N. F. Nasr, Y. E. Elenany, and A. A. M. Hassan, "Isolation and identification of lactic acid bacteria from the intestinal tracts of honey bees, Apis mellifera L., in Egypt", Journal of Apicultural Research, vol. 60, issue 2, pp. 349–357, 2021. 3.pdf
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Elzeini, H. M., A. A. Ali, N. F. Nasr, Y. E. Elenany, and A. A. Hassan, "Isolation and identification of lactic acid bacteria from the intestinal tracts of honey bees, Apis mellifera L., in Egypt", Journal of Apicultural Research,, vol. 60, issue 2, pp. 349-357, 2021.
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Khairy, M., A. Mahmoud, M. H. Sayyouh, A. Osman, and G. A, "Isolation and Identification of Indigenous Bacteria in Some Egyptian Crude Oils", AEIC97, Al_azhar University, Cairo, Egypt, 1997. isolation_identification_and_selection_of_a_suitable_type_of_bacteria_for_meor_in_the_egyptian_oil_fields_-_al-azhar_int.eng_.conf_._-_2004.pdf
Teruaki, A. K. A. O. ‘, and M. HATTORIW, "Isolation and identification of human intestinal bacteria capable of hydrolyzing saikosaponins", Journal of Traditional Medicines, vol. 14, pp. 34-40, 1997. Abstract
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Kilany, W. H., A. M.M, E. - S. A.A, and E. F. El-Khashab, "Isolation and Identification of HPAI from Fertile Egg Derived from Broiler Breeder Farm after Natural Vaccination Breakage during 2010 in Egypt", Animal Health Research Journal, vol. 1, issue 3, pp. 6-16, 2013.
Hamed, W., S. Abdelmalek, and A. E.H.Mostafa, "Isolation and identification of Helicobacter spp. from gastric biopsies and autopsies in human and animal gastric patients", Journal of current Veterinary Research , 2022.
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