EXTENSION OF OUR IMMUNE SYSTEM: RED BLOOD CELLS Swati Mohanty Abstract: Erythrocytes or Red blood cells are

the major apoptotic cells in the physiological system. They are also the most abundant of all the blood cells. For ages these cells have been considered as mere oxygen-carriers. The present hypothesis however is suggestive of varied roles played them in the immune-system. Two attributes which are mainly explored here are eryptosis and membrane-fusibility. Other than conferring innate immunity to the organism, these cells elicit and influence both humoral as well as cell-mediated immunity when targeted or bound by an antigen. They can elicit T-cell depletion and T-cell anergy transiently on targeted binding of antigens on their surface; which can be called as long-lasting or sometimes transient immune-suppression. This process might also lead to an aggravated or sometimes weakened B-cell response (depending upon the T-Cell targeted).There might be occasional hemorrhagic immune-response against antigens (increased complement activity), auto-antibody production and also elimination of auto-reactive self cells. Such antigen binding has already been explored as a drug delivery process and the efficacy of such a system in the treatment of diseases T-cell lymphoma and T-cell leukemia is yet to be established. Keywords: Erythrocyte/RBC, Eryptosis, T-cell depletion, T-cell anergy, transient immunosupression RBC CONTRIBUTES TO INNATE IMMUNE-RESPONSE: It has been recently demonstrated that like neutrophils and endothelial cells, RBC can also synthesize NO(Nitric oxide) through enzymes that use L-Arginine as substrate .Exposure of RBCs to physiological levels of shear stress leads to release of NO which helps to regulate and maintain the vascular tonus. These cells can also produce Hydrogen sulfide gas which relaxes vascular tissues. A major contributer to the cardio-protective nature of garlic are nothing but these tiny cells which convert the sulfur compounds present in them into hydrogen-sulfide. On energy depletion as well as antigenic stress, RBC undergo suicidal or programmed death called eryptosis which is characterized by cell-swelling and phosphatidyl serine exposure at the cell-surface. Even without any inducible factors, eryptosis occurs by caspase 3 and caspase 8 governed pathways. Cation channels also trigger apoptotic death in the RBCs. It is easy to manipulate these cells and turn them into drug-carriers due to their high membrane-fusibility. Here it might easily be hypothesized that these cells have inherent genetic-modulator potential. Lastly these cells can be easily targeted through the surface molecule glycophorin A and this forms the major thrust for the subsequent discussion below.

THE THEORY: Anergy can be defined as the state of lymphoid cells where they do not receive signal for activation to elicit an immune response. Apoptotic cells are a source of tolerogenic antigens ( in this case phosphatidyl serine for RBCs) and the displayed antigen(to the Antigen presenting cells or phagocytes) does not elicit an inflammatory or a strong immune response. This is because the Toll-like receptor signaling is inhibited, T-cells which originally should be activated and migrated to the site turn anergic; else follow rapid deletional proliferation. Here targeted deletion of auto-reactive T-cell type can be achieved with ease as they at first proliferate strongly against RBCs .Again this T-

cell depletion is transient and coherent with the clearance of the eryptotic erythrocyte fraction. Apparent reduction of B-cell activity may occur .But this is reversed if the CD 8+ T-cells are targeted and not CD 4+ T- cells. Thus, targeted erythrocyte and T-cell depletion leads to a transient immuno-suppression where many autoimmune diseases and T-Cell Lymphoma can be treated. Immune -surveillance might differ stating more tumors in individual whose adaptive immune systems are suppressed but this is the case only for strongly immunogenic tumors like skin cancer and would not have any profound effect on non-immunogenic solid tumors or metastases. This theory comes as a respite for the cases where the later can be targeted.

APPLICATION OF THE THEORY TO VARIOUS DISEASES: Previously IL-10 and TGF- are known chemokines which are therapeutically introduced for immune-suppression for auto-immune diseases and organ/protein transplants. The therapy induced Fas-ligand mediated apoptosis. This approach was met with challenges like targeted delivery system and effect on over all physiology. RBCs indeed come as a targeted antigen/peptide carrier. Auto-immune disorders have been successfully treated through targeting RBC in mouse model; a work validated by Stephan et al. (2012).This group successfully demonstrated insertion of two RBC molecule; one with a peptide moiety and the other with a fusion protein carrying an antibody fragment against auto-antigens (both targeting the glycophorin A surface molecule of RBC) in a diabetic mouse model. The results were eryptosis, deletional- proliferation of antigen specific T-cell, lowering of inflammatory response upon immunization and clearance of apoptotic debris. This opens up the arena for also targeting virally infected cells in the case of acute infection, where infected or dysfunctional effector cells are cleared using specific peptide. An example is the case of acute herpes virus infection where Foxp3+ CD4+ T-regs which negatively regulate effective immunity and thus can be targeted for deletional proliferation .In the case of solid tumor cancer, + this theory is also applicable. With the induced CD 8 T cell Clonal anergy, the non-adherent NK (Natural Killer) Cells are at their best at killing the antibody-coated cancer cells through ADCC (Anti-body dependent cellular cytotoxicity). (peptide moiety which can be specific antibody against a T-Cell e.g.Foxp3+ CD4+ antibody+ glycophorin targeting antibody)

Anergic T-cell
eryptosis

T-cells T-cell activation T-cell depletion

Fig: DELETIONAL PROLIFERATION OF T-CELL BY TARGETTING RBC

CONCLUDING REMARKS: A very primary experimentation using a mixture of microbial antigens (coleys toxin) bound to peptide and targeting RBCs shows how the innate immune response can be enhanced with susceptible damage to cancer cells using this toxin and also induction of hemorrhagic necrosis as well as lesions in tumor endothelium without affecting normal vasculature. Furthermore decrease in T-cell number can aid the chemotherapy/organ-transplant in metastatic tumor or T-cell Lymphoma. REFERENCES: 1) Kontos,Stephan, et al.(2012) Engineering Antigens for in-situ erythrocyte binding induces T-cell deletion. PNAS E60-68(December issue) 2) Stermberg,Nadine et al.(2011) Surface Modified loaded human red blood cells for targeting and delivery of drugs.Journ.of.Microencapsulation 1-12 3) Recher,Mike, et al.(2004) Deliberate removal of T-Cell help improves virus-neutralizing antibody prod uction. Nat.immunology Vol.5 No. 9

Swati Mohanty (swatimohanty88@rediffmail.com) MSc microbiology Bhubaneswar