Contributions
Abstract: P759
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Immunology
Background: Neural Precursor Cells (NPCs) of Central Nervous System (CNS) are proposed as cell therapy for neurodegenerative diseases. Despite the inarguable positive results after their transplantation in Experimental Autoimmune Encephalomyelitis (EAE), many issues remain open to question.
Goals: We examined the immune responses after NPCs subcutaneous inoculation in naive mice.
Methods: NPCs were isolated from C57BL/6 neonates and were administered subcutaneously in naive C57BL/6 adult mice. MOG35-55-EAE mice served as a positive control and naive mice as negative control. On day 17 (Acute phase) blood sampling was performed in all experimental groups and corresponding antisera (NPCs-AS, EAE-AS, NAIVE-AS) were collected. Animals were sacrificed for tissue collection on days 17 and 50 (Chronic phase). The antisera were examined for autoantibody presence by Western Blotting on normal spinal cord homogenate and NPCs lysate. Additionally, brain and spinal cord were tested by Real-time PCR (qPCR) for the expression of antigen presentation and inflammation-related genes.
Results: Western Blot revealed dominant immunoreactivity on NPCs substrate indicating specific bands (100kDa, 60kDa, 50kDa, two bands above 40kDa, 30kDa and 25kDa), when using NPCs-AS. On spinal cord substrate one band was yielded (25kDa), suggesting a mere presence of immunoreactivity against CNS. CD80 and CD86 mRNA expression levels were elevated in the brain of NPCs-inoculated group versus EAE-group; CD80-Acute:3.08-fold vs 0.123-fold, p< 0.01, C80-Chronic:3.87 vs 1.69-fold, p< 0.05, CD86-Acute:5.89-fold vs 4.24-fold, CD86-Chronic:3.89-fold vs 0.39-fold, p< 0.01. CCL5 and CXCL1 mRNA were also overexpressed in the brain of NPCs-inoculated group versus EAE-group; CCL5-Acute: 23.94-fold vs 1.31-fold,p< 0.001, CCL5-Chronic:57.53-fold vs 4.24-fold, CXCL1-Acute:8.96-fold vs 4.87-fold; CXCL1-Chronic phase:20.52-fold vs 1.35-fold, p< 0.001. IL-17RA (receptor of IL-17) mRNA was upregulated in spinal cord of both groups in acute phase (NPCs-group:23.21-fold, EAE-group:35.17-fold) and remained increased only in NPCs-group in chronic phase (38.19-fold vs 2.7-fold).
Conclusions: Humoral immunity induced after naive mice immunization with NPCs, provides evidence of NPCs antigenic potential, cells that are considered to be immune privileged. Thus, cross-talk between NPCs and CNS immune mechanisms needs further investigation, before their use as therapeutic agent.
Disclosure: Evangelia Kesidou: Nothing to disclose
Roza Lagoudaki: Nothing to disclose
Olga Touloumi: Nothing to disclose
Kyriaki-Nepheli Poulatsidou: Nothing to disclose
Konstantinos Xanthopoulos: Nothing to disclose
Paschalis Theotokis: Nothing to disclose
Evangelia Nousiopoulou: Nothing to disclose
Evangelia Kofidou: Nothing to disclose
Nickoleta Delivanoglou: Nothing to disclose
Marina Boziki: Nothing to disclose
Ioannis Nikolaidis: Nothing to disclose
Christos Bakirtzis: Nothing to disclose
Georgios Hadjigeorgiou: Nothing to disclose
Constantina Simeonidou: Nothing to disclose
Nikolaos Grigoriadis: Nothing to disclose
Abstract: P759
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Immunology
Background: Neural Precursor Cells (NPCs) of Central Nervous System (CNS) are proposed as cell therapy for neurodegenerative diseases. Despite the inarguable positive results after their transplantation in Experimental Autoimmune Encephalomyelitis (EAE), many issues remain open to question.
Goals: We examined the immune responses after NPCs subcutaneous inoculation in naive mice.
Methods: NPCs were isolated from C57BL/6 neonates and were administered subcutaneously in naive C57BL/6 adult mice. MOG35-55-EAE mice served as a positive control and naive mice as negative control. On day 17 (Acute phase) blood sampling was performed in all experimental groups and corresponding antisera (NPCs-AS, EAE-AS, NAIVE-AS) were collected. Animals were sacrificed for tissue collection on days 17 and 50 (Chronic phase). The antisera were examined for autoantibody presence by Western Blotting on normal spinal cord homogenate and NPCs lysate. Additionally, brain and spinal cord were tested by Real-time PCR (qPCR) for the expression of antigen presentation and inflammation-related genes.
Results: Western Blot revealed dominant immunoreactivity on NPCs substrate indicating specific bands (100kDa, 60kDa, 50kDa, two bands above 40kDa, 30kDa and 25kDa), when using NPCs-AS. On spinal cord substrate one band was yielded (25kDa), suggesting a mere presence of immunoreactivity against CNS. CD80 and CD86 mRNA expression levels were elevated in the brain of NPCs-inoculated group versus EAE-group; CD80-Acute:3.08-fold vs 0.123-fold, p< 0.01, C80-Chronic:3.87 vs 1.69-fold, p< 0.05, CD86-Acute:5.89-fold vs 4.24-fold, CD86-Chronic:3.89-fold vs 0.39-fold, p< 0.01. CCL5 and CXCL1 mRNA were also overexpressed in the brain of NPCs-inoculated group versus EAE-group; CCL5-Acute: 23.94-fold vs 1.31-fold,p< 0.001, CCL5-Chronic:57.53-fold vs 4.24-fold, CXCL1-Acute:8.96-fold vs 4.87-fold; CXCL1-Chronic phase:20.52-fold vs 1.35-fold, p< 0.001. IL-17RA (receptor of IL-17) mRNA was upregulated in spinal cord of both groups in acute phase (NPCs-group:23.21-fold, EAE-group:35.17-fold) and remained increased only in NPCs-group in chronic phase (38.19-fold vs 2.7-fold).
Conclusions: Humoral immunity induced after naive mice immunization with NPCs, provides evidence of NPCs antigenic potential, cells that are considered to be immune privileged. Thus, cross-talk between NPCs and CNS immune mechanisms needs further investigation, before their use as therapeutic agent.
Disclosure: Evangelia Kesidou: Nothing to disclose
Roza Lagoudaki: Nothing to disclose
Olga Touloumi: Nothing to disclose
Kyriaki-Nepheli Poulatsidou: Nothing to disclose
Konstantinos Xanthopoulos: Nothing to disclose
Paschalis Theotokis: Nothing to disclose
Evangelia Nousiopoulou: Nothing to disclose
Evangelia Kofidou: Nothing to disclose
Nickoleta Delivanoglou: Nothing to disclose
Marina Boziki: Nothing to disclose
Ioannis Nikolaidis: Nothing to disclose
Christos Bakirtzis: Nothing to disclose
Georgios Hadjigeorgiou: Nothing to disclose
Constantina Simeonidou: Nothing to disclose
Nikolaos Grigoriadis: Nothing to disclose