HFRS患者特异性IgA、IgE抗体及其免疫复合物测定

Studies on Detection of Serum Specific IgA, IgE Antibodies and Circulation Immune Complexes in Patients of Hemorrhagic Fever with Renal Syndrome

Zhang Donghai
(Zibo Central Hospital. Zibo.Shandong Province 255036)
Sun Hui Gao Feng
(Zibo Second Health School)

Abstract Detection had been made for the levels of serum specificity IgA, IgE antibodie and specificity of virus antigens circulation immune complexes (CIC) on the different days or phases of illness and clinic types in patients of HFRS. The results showed that the level of specific IgA antibody was higher in slight cases than in middle and severe; specific IgE antibody and its CIC was higher in severe cases than in slight and middle. Those differences were apparent in acute phase (illness phase of fever and shock-oilguria, or 3-8 days after illness). But specific CIC IgA types had no diferences.
Key words Hemorrhagic Fever with Renal Syndrome (HFRS), Specific IgA antibody, Specific IgE antibody, Circulating Immune Complexes (CIC), ELISA

对肾综合征出血热(HFRS)免疫发病机理研究较多的是IgG、IgM、IgE型抗体[1～4]。已有报告在HFRS患者血清中可检测到特异性IgA抗体[5～6]。为进一步了解HFRS患者发病过程中特异性抗体及其循环免疫复合物的形成及其致病性，我们平行测定了特异性IgA、IgE抗体以及抗原特异性IgA、IgE型循环免疫复合物(CIC)的水平，并从临床分型、病期、病日等角度作了对比分析。

1 材料与方法

1.1 HFRS病人血清 采集本市54例HFRS病人进出院时血清共108份。IgA型检测108份，IgE型检测其中93份。血清病期、病型分布见表1。

表1 待测血清病期、病型分布
Table 1 Phases and types of illness for collected sera

病期
phase of illness 轻
slight 中
middle 重
severe 合计
total
发热 fever (18) 16 (18) 17 (2) 2 (38) 35
休克少尿 shock & oilguria (1) 0 8 (7) (5) 5 (14) 12
多尿 polyuria (8) 6 (14) 11 (3) 3 (25) 20
恢复 recovery (15) 12 (12 )11 (3) 4 (31) 26
合计 total (42) 34 (52) 46 (14) 13 (108) 93

注： ( )外为IgE型，( )内为IgA型检测血清数。
Note: The numbers out of the parenthesis are the types of IgE antibody tested and in the parenthesis are the numbers of tested IgA.
1.2 试剂

1.2.1 兔抗HFRS血清及鼠脑HFRS抗原 卫生部中国药品生物制品检定所俞永新教授惠赠。
1.2.2 兔抗人IgE抗体购自Dako公司。
1.2.3 鼠抗人IgA抗体及HRP-鼠抗人IgA抗体 系中国医科院基础研究所产品。
1.2.4 HRP-鼠抗HFRS 小鼠腹水系南京军区总院产品，纯化后标记HRP(RZ=3.0)。
1.3 实验方法
1.3.1 检测HFRS-IgA、IgE抗体 采用改良抗体捕获ELISA技术[5]。即包被抗人IgA或IgE-McAb→病人血清→HFRSV-Ag+HRP-抗HFRS。
1.3.2 检测IgA型CIC 采用双抗体夹心法(另文报告)。即包被兔抗HRFS→病人血清(HFRSV-IgA抗体)→HRP抗人IgA。
1.3.3 检测IgE型CIC 采用捕获法。参见文献[7]。即包被兔抗人IgE→病人血清(IgE抗体-HFRSV)→HRP-抗HFRS。

2 结果

2.1 不同病期、临床型HFRS病人血清中特异性抗体及特异性CIC水平(几何平均滴度、GMT)及阳性率结果。见表2。

表2 不同病期HFRS病人血清特异性IgA、IgE抗体及其特异性
CIC的GMT值与阳性率测定结果
Table 2 The results of specific IgA, IgE antibodies and specific circulating immue complex in different illness phases of HFRS

抗体型别
type of
antibody 病期
phase of
illness 病例数
numbers of case 轻型slight (positive rate)(%) 中型middle (positive rate)(%)
抗体specific
antibody CIC antigen specific 抗体specific
antibody CIC antigen specific
发热fever 38 104.76(100.00) 11.41(55.56) 34.34(83.33) 3.21(61.11)
休克少尿 14 320.00(1/1) 80.00(1/1) 95.14(100.00) 17.84(87.50)
shock & oilguria
IgA型 多尿polyuria 25 246.75(100.00) 21.87(62.50) 185.62(100.00) 30.25(85.71)
恢复recovery 31 367.58(100.00) 28.99(80.00) 213.58(100.00) 11.48(58.33)
综算total 108 204.88(100.00) 18.88(66.67) 96.43(94.23) 10.25(63.46)

发热fever 35 29.58(81.25) 11.76(56.25) 57.81(88.24) 15.28(70.59)
休克少尿 12 - - 114.95(100.00) 12.63(71.43)
shock & oilguria
IgE型 多尿polyuria 20 100.80(100.00) 7.10(50.00) 249.99(100.00) 15.26(63.64)
恢复recovery 26 269.12(100.00) 5.53(41.67) 386.53(100.00) 7.48(54.55)
综算total 93 80.06(91.18) 8.24(50.00) 148.64(95.65) 12.51(67.39)
抗体型别
type of
antibody 病期
phase of
illness 病例数
numbers of
case 重型severe (positive rate)(%) 合计total (positive rate)(%)
抗体specific
antibody CIC
antigen specific 抗体specific
antibody CIC
antigen specific
发热fever 38 56.57(2/2) 0.00(0/2) 59.79(92.11) 5.51(44.74)
休克少尿 14 28.99(80.00) 43.94(80.00) 67.87(92.86) 27.40(85.71)
shock & oilguria
IgA型 多尿polyuria 25 507.97(100.00) 40.00(100.00) 229.44(100.00) 28.20(80.00)
恢复recovery 31 320.00(100.00) 26.75(75.00) 292.62(100.00) 20.05(70.93)
综算total 108 116.96(92.86) 21.76(71.43) 132.54(96.30) 14.33(65.74)

发热fever 35 160.00(2/2) 56.57(2/2) 45.11(85.71) 14.61(65/71)
休克少尿 12 160.00(100.00) 69.64(100.00) 151.04(100.00) 25.72(83.33)
shock & oilguria
IgE型 多尿polyuria 20 507.97(100.00) 7.37(66.67) 211.72(100.00) 10.88(65.00)
恢复recovery 26 640.00(100.00) 9.28(66.67) 346.64(100.00) 6.67(50.00)
综算total 93 287.63(100.00) 45.37(84.62) 130.01(94.61) 11.85(63.44)

注：( )外数字为GMT值，( )内数字为检出率。
Note: The numbers out of the parenthesis are the values of GMT (Geometric Mean Titer) and the numbers in the parenthesis are the positive rate.
2.2 不同病日检出结果。见表3。

表3 不同病日HFRS患者血清特异性IgA、IgE抗体及其特异性
CIC的GMT值与检出率测定结果
Table 3 The results of specific IgA, IgE antibodies and specific circulating immune complex on different days of illness in HFRS patients

抗体型别
type of
antibody 病日
days of
illness 病例数
numbers of
case 轻型slight (positive rate)(%) 中型middle (positive rate)(%)
抗体specific
antibody CIC
antigen specific 抗体specific
antibody CIC
antigen specific
0～ 10 126.96(100.00) 5.63(33.33) 11.70(2/3) 7.37(1/3)
4～ 40 104.39(100.00) 18.36(61.54) 50.98(95.24) 4.60(47.62)
IgA 8～ 21 319.89(100.00) 26.75(75.00) 174.44(100.00) 11.93(62.50)
型 12～ 19 289.75(100.00) 22.87(71.43) 170.36(100.00) 23.20(81.82)
16～ 11 320.00(2/2) 160.00(2/2) 201.57(100.00) 24.28(83.33)
20～28 7 905.10(2/2) 28.28(2/2) 403.17(100.00) 6.84(33.33)
综算total 108 204.08(100.00) 18.88(66.67) 96.47(94.23) 10.05(63.46)

0～ 10 27.25(83.33) 14.74(66.67) 23.39(2/3) 40.00(2/3)
4～ 36 36.06(81.82) 15.26(63.64) 84.14(94.74) 15.15(73.68)
IgE 8～ 15 148.14(100.00) 7.01(44.44) 228.03(100.00) 7.61(40.00)
型 12～ 18 226.40(100.00) 2.08(16.67) 319.92(100.00) 18.82(72.73)
16～ 8 320.00(1/1) 40.00(1/1) 320.00(100.00) 6.93(60.00)
20～28 6 640.00(1/1) 0.00(0/1) 285.08(100.00) 2.71(33.33)
综算total 93 80.06(91.18) 8.24(50.00) 148.64(95.65) 12.51(67.39)
抗体型别
type of
antibody 病期
days of
illness 病例数
numbers of
case 重型severe (positive rate)(%) 合计total (positive rate)(%)
抗体specific
antibody CIC
antigen specific 抗体specific
antibody CIC
antigen specific
0～ 10 80.00(1/1) 0.00(0/1) 59.29(90.00) 3.81(40.00)
4～ 40 30.60(83.33) 23.39(66.67) 59.61(92.50) 9.20(57.50)
IgA 8～ 21 1280.00(1/1) 160.00(1/1) 271.24(10.00) 21.42(71.43)
型 12～ 19 40.00(1/1) 40.00(1/1) 191.97(100.00) 23.75(78.95)
16～ 11 403.20(3/3) 18.57(2/3) 268.47(100.00) 31.79(81.82)
20～28 7 640.00(2/2) 28.28(2/2) 579.66(100.00) 15.39(71.43)
综算total 108 116.96(92.86) 21.76(71.43) 132.85(96.30) 14.33(65.74)

0～ 10 40.00(1/1) 20.00(1/1) 27.05(80.00) 20.50(80.00)
4～ 36 201.58(100.00) 80.00(100.00) 75.13(91.67) 20.04(75.00)
IgE 8～ 15 160.00(1/1) 0.00(0/1) 171.93(100.00) 6.33(40.00)
型 12～ 18 640.00(1/1) 20.00(1/1) 296.29(100.00) 9.06(55.56)
16～ 8 639.68(2/2) 4.47(1/2) 380.54(100.00) 7.73(62.50)
20～28 6 905.10(2/2) 28.28(2/2) 479.45(100.00) 5.02(50.00)
综算total 93 287.63(100.00) 45.37(84.62) 130.01(94.62) 11.85(63.44)

注：同表2。 Note: as Table 2.
3 讨论