Journal of Babylon University/Pure and Applied Sciences/ No.(1)/ Vol.(22): 2012

College of Science/Babylon University Scientific Conference

Estimationof the concentrations of complement components , alph-1-antitrypsin and alph-2 macroglobulin in patients with B- thalassemia

Alaa J.H. ; Shakir , H.M. ; Zina , H.M. ; Raheem , T.O. , and Mohamed , H.A.

1,2,5 University of Babylon , College of Science , Biology Department

3, 4 Babylon Health Directorate

Abstract :

An immunological study were performed to estimation the concentrations of complement components ( C3 and C4), alph-1 antitrypsin and alph-2 macroglobulin in sera of patients with B - thalassemia in Babylon province . The concentrations of these immunological parameters were determined by single radial immunodiffusion assay (SRID) . This study revealed decreases in levels of complement components ( C3 and C4) in patients which reached 89.2 and 19.9 mg/dl respectively , while it was 96.7 and 21.9 in control group , furtheremore, this study revealed decreases in levels of alph-1 antitrypsin in patients in comparison with the healthy subjects , while there was no differences in mean of concentrations of alph-2 macroglobulin between these two groups . The results illustrated that decreases in levels of complement components as well as alph-1 antitrypsin in patients compared with healthy subjects in babylon province.

keywords: Complement . Alph-1 antitrypsin . Alph-2 macroglobulins . Patients . B- Thalassemia.

Introduction :

The increased frequency of infections associated with B- thalassemia major seems to be related to abnormalities of the immune system ( Lombardi et al. ,1994 ; Uguccioni et al. , 1993 ) , and the predisposition to autoimmune diseases is under the control of immune response genes , which play a central role in the presentation of antigens to the immune system ( Bottazzo et al. , 1986) . B- thalassemia major itself is not a true hemochromotosis , it is a secondary hemochromatosis , which is based on the combination of chronic hemolytic anemia , iron intestinal hyperabsorption, and multiple blood transfusions that the patients receive ( Jessup and Manno , 1998 ) . Similarities between microbial antigens and self HLA molecules may result in autoimmune reaction after infections (Abbas et al. , 1997) . B- thalassemia is caused by defect of synthesis the beta- globulin chains of the hemoglobin tetramer ( Cao , 2010 ; Luca et al. , 2010 ) , as a result the patients suffer from chronic anemia due to a process called ineffective erythropoiesis with massive splenomegaly and dramatic iron overload ( Libani et al. , 2008 ; Luca et al.,2001). The serum complement levels were found in some studies to be normal ( Vergin et al. , 1997 ;Corry et al. , 1981 ) , and in some patients a deficiency in the hemolytic activity of the classical(ConstantouLakis et al. 1978 ) or the alternative complement pathways was detected( Corry et al. , 1981 ) . Romeo et al. (1983) investigated a possible relationship between alph-1 antitrypsin phenotypes and liver disease in transfusion dependent thalassemia subjects . Alpha -2 macroglobulin is a large plasma protein found in the blood , and it is produced by the liver , as well as is amajor component of the alpha -2 band in protein electrophoresis , also this protein is able to inactivate an enormus variety of proteinases ( including serine , cysteine , aspartic and metalloproteinases ) , and it functions an inhibitor of coagulation by inhibiting thrombin , and inhibitor for fibrinolysis by inhibiting plasmin and kallikrein ( Do Boer et al. , 1993 ; Anderson et al . 1995)

The aim of this study to evaluate the changes in some humoral parameters of immune system in B- thalassemia patients in babylon governorate .

Materials and Methods :

1- Patients and Control :

Twenty five patients ( 15 males and 10 femles ) with B- Thalassemia their agesranged berween 10 - 20 years were studied at the thalassemic center in maternity and pediatrics hospital ( Babylon province ) . All patients were living in babylon province and pre- diagnosed by physician. Fifteenth apparently healthy subjects (8 males and 7 females) with no symptoms of thalassemia disease were selected as control group .

2- Blood Samples :

The blood samples were drawn from each patients and controls (5 ml) by vein puncture using disposible syringes . The blood was placed in disposible tube , kept to clot at room temperature , and then centrifuged at 3000 r.p.m ( Bishop et al , 1985) for 10 minutes . Sera were carefully transferred to appendrof tubes and stored in a liquotes at deep freezing at -20 C until used .

3- Immunological Tests :

The concentrations of complement components ( C3 and C4), al ph-1 antitrypsin and alph-2 macroglobulin were estimated according to Mancini et al , (1965) to which and manual procedure of linear chemical procedure to which( company of Al macen Joaquim . Costa, Motgat , Barcelona , Spain

4- Statistical Analysis :

The results were analyzed using completely randomized design (Donald, 1989).

Results:

The means of complement components C3 and C4 in patients were lower compared with control group , which reached 89.2 and 19.9 mg/dl respectively , while it was 96.7 and 21.9 mg/dl consuctively in control group as illustrated intable (1)

Table (1) : The concentrations of complement components C3 and C4 (mg/dl) in patients with B- thalassemia .

Parameter
Nature
Of treatment / Conc. Of C3 (mg/dl)
Mean -+ SE / Conc. Of C4 (mg/dl)
Mean -+ SE
Patients / 89.2-+ 7.4 / 19.9-+2018
Control / 96.7 -+ 8.6 / 21.9 -+ 2.8

Meantime,the means of concentrations of alph-1 antitrypsin (mg / dl) were slightly lower in patients as compared with control group, while there was no significant difference in concentration of alph-2 macroglobulin between patients and control group as revealed in table (2)

Table (2) : The concentrations of alph-1 antitrypsin and alph-2 macroglobulin (mg/ dl) in patients with B- thalassemia .

Parameter
Nature
of treatment / Conc.of alph-1 antitrypsin (mg/dl)
Mean -+ SE / Conc. of alph-2 macroglobulin (mg/dl)
Mean -+ SE
Patients / 112.5 -+ 10.8 / 157.5 -+ 12.8
Control / 120.2 -+ 10.5 / 158.6 -+ 16.2

Discussion :

The patients with B- thalassemia showed lower values of serum complement components C3 and C4 . The donation is responsible for autoantibodies to the antigens involved which are derived from the donars because almost are HLA incompatibilities . The component of the immune system includin natural killer cells , were so depressed in multiple transfusion as it happened in aquired immunodeficiency virus infected people ( Pedro et al. , 1992) , also Rund and Rachmilewitz (2005) reported that immunological abnormalities in patients with B- thalassemia included : decreased opsonization and granulocyte phagocytosis as well as alteration in B and T cell number and function. ( Kapadia et al. , 1980 ; Munn et al. , 1981) . Factors such as splenectomy , iron overload ( Kapadia et al. , 1980 ) , repeated exposure to foreign antigens at the time of blood transfusion ( Gascon and Young , 1984) , and the use of the chelating agent ( feferoxamine ) ( Lederman et al. , 1984 ) have profound effects on the immune system . Iron overload has been a major cause of immunological disturbance in thalassemia , the mechanisms suggested include toxic effects of high iron levels on lymphocyte function ( Bryan and leech , 1981 ) . Ahmed et al.,(2005) illustrated that serum levels of complement componeuts C3 and C4 were lower in patients with B- thalassemia in comparison with healthy subjects . Splenectomized patients had no differences in values of C3 and C4 than non-splenectomized patients . Serum complement components C3 and C3 were consistently reduced in our patients , and this can be either due to reduced synthesis or increased consumption ; the latter is more probable with the rate of infection in patients which seems to be high . Our observation of significantly lower C3 and C4 levels early in life in thalassemic patients , however , points to the possibility of deficient complement synthesis may also act as a contributing factor to immune deficiency . The controversy concerning alteration of serum complement levels in B- thalassemia patients may be due to marked heterogeneity of the patients in different studies , and this heterogeneity concerns race , socioeconomic class , nutritional status and environmental factors ( Vergin et al. , 1997 ) .

The table(2) results revealed increases in alph -1 antitrypsin levels in B-thalassemic patients , while there is no differences between patients and control groups in values of alph-2 macroglobulin . Romeo et al. , (1983) illustrated that the mean concentration of alph-1 antitrypsin was significantly increased in thalassemic patients . Acommon variant ( polymorphism ) , of alph-2 macroglobulin leads to increased risk of Al zheimer's disease , also its levels are increased in nephritic syndrome ( Blacker etal ., 1998 ; Kavacs , 2000) . Individuals inheriting two B- thalassemic alleles experience deficit in beta – chain production , which lead to excess production of alph- globulin ( Luca etal. , 2010) . Furtheremore , excess alpha – globulin chains in B- thalassemia is creating tetramers of hemoglobin that accumulate and precipitate in the erythroid progenitors , forming inclusion bodies that cause exidative membrane damage within the RBCs and immature developing erythroblasts in the bone marrow ( Libani etal. , 2008) .

Conclusions :

These results can be due to continous exposure to antigens , repeated infections , chronic liver disease and splenectomy . The only probable cause of humoral immune deficiency found in these patients is a defect in serum complement levels , while there is an increases in alph-1 antitrypsin level.

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