在低离子强度溶液（LISS）存在下吸附自身抗体，以检测温自身抗体掩盖的同种抗体。

Chiaroni Jacques, Touinssi Mohammed, Mazet Magali, De Micco Philippe, Ferrera Virginie

Etablissement Français du Sang Alpes Méditerranée, 149, Boulevard Baille, 13005 Marseille, France.

Transfusion. 2003 May;43(5):651-5. doi: 10.1046/j.1537-2995.2003.00356.x.

BACKGROUND

The safe transfusion of patients with warm autoimmune hemolytic anemia requires an efficient and time-saving assay to detect alloantibodies underlying autoantibodies. Methods used include RBCs treated with ZZAP reagent, proteolytic enzyme, or untreated RBCs in the presence of PEG. We propose a method using LISS, which presents some advantages over previous methods.

STUDY DESIGN AND METHODS

We evaluated the effectiveness of autoantibody adsorption with papain-treated and untreated RBCs in the presence of LISS for removal of autoantibodies, without affecting alloantibodies.

RESULTS

One-hundred twenty sera containing autoantibodies were adsorbed with our routine method, which uses papain-treated allogeneic RBCs. Seven-hundred twenty adsorptions (mean, 6 per sample) and 21,600 minutes (mean, 180 min per sample) were required to remove autoantibodies. Fifty sera were adsorbed with our routine method that uses papain-treated allogeneic RBCs in the presence of LISS. The number of adsorptions and the completion time were, respectively, 144 (mean, 2.9 per sample) and 2,880 minutes (mean, 57.6 min per sample). Twenty sera were evaluated with untreated autologous RBCs, in the presence of LISS; 58 adsorptions (mean, 2.9) and 1,160 minutes (mean, 58 min per sample) were required. Three adsorptions with antigen-negative allogeneic RBCs were performed on 8 sera containing alloantibodies with weak reactivity (anti-K [1], anti-D [1], anti-Fya[2], anti-S [2], anti-E [1], and anti-Jka[1]). Alloantibodies were detected with the LISS procedure with papain-treated and untreated RBCs and the routine papain method. Alloantibodies with weak reactivity, tested against the same antibody-detection RBCs, remained unchanged following adsorption of 5 sera. An anti-S, with very weak reactivity, was no longer detected, and an anti-Jka became weaker (1+), regardless of the procedure used. One anti-D became weaker only with the LISS adsorption method.

CONCLUSION

Autoantibodies can be adsorbed more efficiently in the presence of LISS.

背景

对于温抗体型自身免疫性溶血性贫血患者进行安全输血，需要一种高效且节省时间的检测方法来检测自身抗体掩盖下的同种抗体。所使用的方法包括用ZZAP试剂处理的红细胞、蛋白水解酶处理的红细胞，或在聚乙二醇存在下未处理的红细胞。我们提出一种使用低离子强度盐溶液（LISS）的方法，该方法比以前的方法具有一些优势。

研究设计与方法

我们评估了在LISS存在下，用木瓜蛋白酶处理和未处理的红细胞吸附自身抗体以去除自身抗体而不影响同种抗体的有效性。

结果

采用我们的常规方法（使用木瓜蛋白酶处理的同种异体红细胞）对120份含有自身抗体的血清进行吸附。去除自身抗体需要720次吸附（平均每份样品6次）和21600分钟（平均每份样品180分钟）。采用我们的常规方法（在LISS存在下使用木瓜蛋白酶处理的同种异体红细胞）对50份血清进行吸附。吸附次数和完成时间分别为144次（平均每份样品2.9次）和2880分钟（平均每份样品57.6分钟）。使用未处理的自身红细胞在LISS存在下对20份血清进行评估；需要58次吸附（平均2.9次）和1160分钟（平均每份样品58分钟）。对8份含有弱反应性同种抗体（抗-K [1]、抗-D [1]、抗-Fya[2]、抗-S [2]、抗-E [1]和抗-Jka[1]）的血清，用抗原阴性的同种异体红细胞进行3次吸附。采用LISS程序，用木瓜蛋白酶处理和未处理的红细胞以及常规木瓜蛋白酶方法检测同种抗体。用相同的抗体检测红细胞检测，5份血清吸附后，弱反应性同种抗体保持不变。一种反应性非常弱的抗-S不再被检测到，抗-Jka变弱（1+），无论采用何种程序。一种抗-D仅在LISS吸附方法下变弱。