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

  3.1 特异性抗体测定

  据检测结果可见,无论是轻、中或重型病例,HFRS-IgA,IgE抗体检出水平(GMT值)均随病期(病日)进展而逐步升高,这符合一般抗体产生规律。但从两型特异性抗体检出率角度分析,则上述差异不明显。

  HFRS-IgA抗体GMT值在轻型病例高于中、重型病例(P<0.05),这一差异在发热期到休克少尿期(或是8病日以前)表现得尤为突出,至恢复期(或16病日后)则表现不明显。HFRS-IgE抗体GMT值在重型病例始终高于中、轻型,其差异在发热期或病程第一周尤为明显(P<0.05),支持该抗体在病程早期参与病理损伤的观点[4,8]。

  3.2 抗原特异性CIC测定

  IgA型特异性CIC的检出率及GMT值在病型间均无明显差异(P>0.05)。从病期进展或病日角度看,全部病例的检出率及GMT值均以休克少尿及多尿期为高,以发热期为低;但在休克少尿、多尿及恢复期间均无明显统计学差异(P>0.05)。

  IgE型特异性CIC的检出率及GMT值则在轻、中型与重型病例间存在明显的差异(P<0.05),均以重型病例为高。若从病期进展角度看,则轻、中型病例在各病期间无明显差异,但重型病例在发热与休克少尿期明显高于多尿与恢复期(P<0.05)。从病日角度看,则见IgE型特异性CIC的检出率及GMT值均以0~7病日最高,其差异性在重型病例表现得尤为突出。

  上述研究结果显示: HFRS-IgA型抗体在病程早期即可较明显增高,且增高水平以轻型病例为最高。而HFRS病毒特异性IgA型CIC在病型、病日、病期间差异则不显著,水平较低。HFRS-IgE型抗体及其CIC检出率和检出水平均为重型病例明显高于轻、中型病例。因此,对特异性的HFRS-IgA,IgE型抗体及其CIC在参与HFRS免疫损伤的意义值得进一步探讨。

  作者单位:张东海(淄博市中心医院,山东淄博 255036)

  孙 辉 高 峰(淄博第二卫生学校,山东淄博 255015)

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  原作者: 张东海 孙 辉 高 峰