Alterations of red blood cell immunoadherence function in hepatitis B patients

Zi-Qin Sun, Yao Jun Wang, Qi Zhen Quan, Rui Ming Xiao, Feng Guo

Subject headings hepatitis, viral, human/immunology; erythrocytosis/immunology; autigen-antibody complex/blood

Sun ZQ, Wang YJ, Quan QZ, Xiao RM, Guo F. Alterations of red blood cell immunoadherence function in hepatitis B patients. China Nati J New Gastroenterol,1996;2(1):20-21

Abstract

AIM  To investigate the alterations of RBC immunoadherence function in patients with various hepatitis B.

METHODS  RBCC3bRR, RBCICRR and serum CIC levels were measured in 42patients with acute and chronic hepatitis B at active and convalescence stages. 

RESULTS  RBCC3bRRs at the active/acute stage of various hepatitis  were decreased.  They were 13.54%  5.23% in AH, 7.61%  4.12% in AFH,and 16.18%  6.10% in CH,  respectively, all of which were lower than those in normal persons(18.12%  3.91%). At the  quiescent/recovery  stage  of various hepatitis, the RBCC3bRRs were  increased  significantly.  The changes of RBCICRR  and serum CIC level  were contrary to those of RBCC3bRR.

CONCLUSION  RBC immunoadherence function is decreased in acute and chronic  hepatitis. The decrease is in direct proportion to the severity of the diseases.



INTRODUCTION
Red blood cell(RBC) has a lot of immunological functions. The most  important  one is its immunoadherence function, to which much attention  has been paid in recent  years.In previous studies^£Û1,2£Ý, we observed that RBC immunoad herence function in rats with acute  liver  damage  induced by D-galactosamine was  significantly depressed. In the present study, RBC immunoadherence function was detected in patients  with  acute  and chronic hepatitis B by means of the RBC C3b receptor yeast rosette rate(RBCC3bRR) and the RBC immune  complexes rosette rate(RBCICRR) test.

PATIENTS and METHODS

Patients
Forty-two patients with acute and chronic hepatitis B  were chosen in the study. The diagnosis was made according to the diagnostic criteria for hepatitis revised by the National Viral Hepatitis Conference held in Shanghai in 1990. Among them  were  14 patients with acute hepatitis(AH, 9 males and 5 females, mean age 39.5 years), 7 patients  with acute fulminent hepatitis(AFH, 5 males  and 2 females, mean age  35.4  years),  21 patients with chronic  hepatitis(CH,  17 males and 4 females, mean age  36.3 years).All  patients  had evidence of HBV infection. The blood specimens were taken from  the  patients at the  active(or acute)stage and the convalescent stage. Fifteen cases of blood donors were chosen  as  normal controls(NC, mean age 35.6 years).ª¤ª¤ 

Methodsª¤

RBCC3bRR test
One mL of heparin-anticoagulated blood specimen was mixed with Fiscol liquor and was centrifuged to separate red blood cells. The red  blood  cells were washedthree times with normal saline and resuspended in it(1.25 10  cells/ml). Then 0.5ml of complement-sensitized yeast(1¡Á10/ml) was added  into  0.5 ml of the RBC suspension, which was incubated at 37¡æ in a constant-temperature water bath for 30 min. After being diluted with 1ml of normal saline, the RBC was fixed by 0.3?ml of  0.25% glutaraldehyde. Alittle of the prepared RBC suspension was smeared on slide  and  stained with Wright's stain. The number of RBC combining with two yeasts or more was calculated  among 200 RBCs and figured out the RBCC3bRR.ª¥ª¤ 

RBCICRR test
This procedure  was  the same as the above described  except  that the yeast used  was not  sensitized by complement. Circulating immune complexes(CIC) test. Serum CIC was detected  by polythylene glycol precipitation method.ª¤ª¤ 

RESULTS

RBCC3bRR change in various hepatitis
The RBCC3bRRs at the active stage of acute and chronic hepatitis  were decreased. The more severe the disease, the lower  the  RBCC3bRR.  There  was no significant difference between AH  and CH. The RBCC3bRR was increased gradually when the patients recovered. Of the 7 patients with AFH, 3 died of acute liver failure. RBCC3bRRs  in the four survivors were also significantly increased. In CH group,however, the increase was much slower.ª¤ª¤

RBCICRR changes in various hepatitis
In various hepatitis, the RBCICRRs were higher than those in normal  controls(in  the  order of AFH£¾CH£¾AH£¾NC), which was inversely proportional to RBCC3bRR changes(r=-0.863, P£¼0.05). ª¥ª¤

Serum CIC level
Serum level of  immune complexes was higher in patients with hepatitis than that in  individuals,  which was correlated  strongly  with RBCICRR changes(r=0.824, P£¼0.05), but negatively with RBCC3bRR changes(r=-0.959, P£¼0.01).ª¤ª¤

Table 1 Changes of RBCC3bRR, RBCICRR and serum CIC in various hepatitis

A=active; C=convalescentª¤

DISCUSSION
The present study demonstrated that RBCC3bRRs in acute and  chronic hepatitis were decreased and the degree of decrease was closely related to the severity of liver damage. When the patients were recovered or in quiescent stage, their RBCC3bRR became elevated. Therefore, RBCC3bRRs can be used to assess the severity and  prognosis of various hepatitis.ª¥
        It has been verified that there are type I complement receptors (CR1) on the surface of red blood cells, through which red blood cells combine with C3b-opsonized  immune  complexes,  and bring them to macrophages in liver and  other organs,  and finally are eliminated^£Û3,4£Ý. RBCC3bRR and RBCICRR  tests  are  reliable and convenient for detecting RBC immunoadherence function^£Û5£Ý. The decrease of  RBCC3bRR  suggests that the  number  of CR1 is reduced and its function is diminished. In a recent in vivo study^£Û1,6£Ý, we observed that red blood cell immunoadherence  function  in  acute liver  damage was depressed significantly. The degree of the depression was strongly related to the increase of hepatic and serum lipid peroxidate(LPO) level. The ª«in vitroª« study also showed that LPO could  reduce RBCC3bRRs. Antioxidant therapy with vitamin E could resist the depression. The reason  may be  that LPO can combine with amino acid of CR1 protein, causing intra- and inter-cross linking, and thus destroy the normal structure of CR1, which  leads  to the decrease of RBC immunoadherence function.ª¥
        In the present study, we found  that the circulating immune complex wasincreased,  which was related to the decrease of RBCC3bRR. Because CR1 plays an  important  role in  eliminating CIC,  and about 95% of total C3b receptors  in the circulation are on the RBC surface,  the increase of  RBCICRR  reflects  in part the increase of CIC. The mechanism of the decrease of RBC immunoadherence function may be a complicated one.  Further study will be beneficial to the prevention and treatment of hepatitis.

REFERENCES

1  Sun ZQ,  Wang YJ, He JW, Guo F. The dynamic  changes  of red  blood cell immunoadherence function and lipid  peroxide  in
    acute liver failure in rats. Shanghai J Immunol, 1993;13(4): 231-232
2  Sun ZQ, Wang YJ. The coherence between red blood cell immuno-adherence function and lipid peroxidate in acute liver
    damage.   Chin J Exp Clin Immunol, l995;7(1):29-31
3  Siegel I. The red cell immune system. Lancet, 1981;2(8246):556-557
4  Sun ZQ, Wang YJ. Changes of RBCIA and its machanism in acute liver damage. Chin J Clin Hepatol, 1993;9(Suppl):65-66
5  Guo F, Yu ZQ, Zhao ZP. Preliminary study on the  immune  function of human red cells. Chin J Intern Med,
   1982;62(12):715-716
6  Sun ZQ, Wang YJ, Quan QZ, Zhang ZJ. Dynamic changes of RBC  immunoadherent function and lipid peroxidation and  effect of
    vitamin E in acute hepatic injury. Chin J New Gastroenterol, 1996;4(1):6-8

^£±Dr. SUN Zi-Qin, Associate Chief Physician, Department of Gastroenterology,General Hospital of Chinese PLA Jinan Commanding Area, Jinan 250031 Shandong Province China. ª¤
^£±Dr. WANG Yao-Jun, Physician-in-Charge.
^£±Dr. QUAN Qi-Zhen, Chief physician.
^£²Dr. XIAO Rui-Ming, Professor of infectious diseases, Changhai Hospital, Second Military Medical University,
Shanghai 200433 China.
^£²Dr. GUO Feng, Professor of Immunology.ª¤
Tel. +86¡¤531¡¤5952171.
Received  30 October 1995, revised14 November 1995.