**6. Conclusions**

All the previous work in RBC proteomics neither has identified another function nor has mentioned the finding of: HLA, TSAs, or foreign proteins. The reasons are obvious. Firstly, it is not expected to find such proteins and consequently the method used for the interpretation of the mass spectrometry data, and the search engines used for the identification do not consider the right types of sequence data banks available. Secondly, the amount of most of the antigens which belong to the RBC antigens' store is little. This makes those antigens invisible and hence easily missed.

The work described is just a pilot study that throws some light on a new theory related to RBC. This theory is based on finding antigens' store consisting of self and non-self antigens. Although this theory can be related to immune tolerance by logical induction, the concrete evidence and mechanism need further research. Mainly, the logical induction is based on finding all kind of antigens in hemolysate, especially HLA antigens which are related to fetus. This existence of all kinds of antigens, definitely, plays some immunological role which may be immune tolerance.

The initial experiment, which shows the existence of ABO antigens in hemolysate of pregnant females, explains the mechanism of how HDFN occurs. Meanwhile, the

experiment which shows that HLA antigens exist in their hemolysates proposes a new mechanism by which a pregnant woman is able to tolerate her fetus and placenta. Simply, it is the same mechanism a body tolerates his self antigens.

Rediscovering Red Blood Cells: Revealing Their Dynamic Antigens Store and Its Role in Health and Disease 29

developed a computer program that implements the mathematical model to help in its

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The experiments which use hemolysate against self-serum: Ouchterlony immuneprecipitation test, Western Blot, and 2-DE of co-immunoprecipitated antigens demonstrated that RBC have an antigens' store. Mass spectrometry of spots obtained from 2-DE gel demonstrated the finding of all kind of antigens, self and non-self, in hemolysate. This indicates that blood circulating antibodies in any individual will react with his RBC's hemolysate antigens. In effect, there is no absolute immune response, too.

This directed our attention to use hyper immune serum against *Mycobacterium* antigens. This will help to get rid of other proteins and do better separation; and hence better identification. Consequently, we could identify 11 proteins from 60 gel spots belonging to H37Rv strain. The rest of spots are proteins related to bacterial commensals. Consequently, purification of specific antibodies from hyper immune serum is recommended to get further better separation.

In the experiment which investigates the dynamics of foreign antigens in RBC using sheep RBC which has been infected with *E. coli*, it was shown that the concentration of foreign antigens in RBC varies by time in relation to IR known behavior. This proves that RBC have role in immune reactions (IR and IT).

Whatever the reason of this existence of antigens in hemolysate this existence can help in designing diagnostic kits for different types of diseases. Further, it will help in discovering, not only, new immunological disorders which are, now, categorized under idiopathic disease, but also, identifying the obscure cause of many immunological disorders, including cancer. The identification of the cause of a disorder will help in its treatment and prevention.
