Allergological studies on bronchial asthma due to insects - crossreactivity and unique antigenic fraction of insects

1. 利用RAST抑制试验和针对上述抗原的IgE抗体的季节性波动规律研究家蚕翅、蛾蝇翅、摇蝇全身和螨提取物的抗原交叉反应性。1)相互RAST抑制试验表明，在高抗原浓度下，蚕翅、蛾翅和摇蚊全身提取物之间几乎存在完全的交叉反应。但是，这些昆虫与螨之间没有交叉反应，表明昆虫引起的过敏与螨的过敏是独立的。2)针对这些昆虫的IgE抗体的季节波动模式并不平行。这可能意味着这些昆虫在临床水平上的交叉反应性较低。这些相互矛盾的结果促使我们从分子基础上研究抗原部分。提取物的SDS-PAGE分离和独特抗原部分的免疫染色分析。昆虫提取物经2-巯基乙醇还原，SDS-PAGE分离后，检测到至少16条家蚕翅膀的条带，7条家蚕翅膀的条带和7条手蛾全身的条带，其中75-79K、67-68K、63-64K、40-42K、32-33K和28-30K道尔顿反应为家蚕翅膀特异性IgE抗体，67-68K、49-50K和30-31K道尔顿反应为家蚕翅膀特异性IgE抗体。40-42K和25-27K道尔顿对chironomid全身特异性IgE抗体有反应，在超过一半的哮喘患者中。这些结果表明，有必要进一步从分子基础上研究昆虫免疫反应带之间的交叉反应性。

1. Studies on antigenic cross-reactivity between extracts of silkworm wing, caddis fly wing, chironomid whole body, and mite, by use of RAST inhibition tests and seasonal fluctuation patterns of IgE antibodies against the above-mentioned antigens. 1) Mutual RAST inhibition tests showed that at high antigenic concentrations, there were almost complete cross-reactibities among silkworm wing, caddis fly wing, and chironomid whole body extracts. But, there was no cross-reactivity between these insects and mite, showing that the allergies due to the insects were independent from mite allergy. 2) Seasonal fluctuation patterns of the IgE antibodies against these insects did not run parallel with each other. This might mean the less cross-reactivity among these insects at clinical levels. These conflicting results promote us to study the antigenic fractions at the molecular basis.2. SDS-PAGE fractionations of the extracts and immunostaining analyses of the unique antigenic fractions. When the insect extracts were reduced by 2-mercaptoethanol and fractionated by SDS-PAGE, at least 16 different bands for silkworm wing, 7 bands for both caddis fly wing and 7 bands for chironomid whole body were detected, of which 75-79K, 67-68K, 63-64K, 40-42K, 32-33K, and 28-30K daltons were reacted to silkworm wing-specific IgE antibodies, 67-68K, 49-50K, and 30-31K daltons were reacted caddis fly-specific IgE antibodies, and 40-42K and 25-27K daltons were reacted to chironomid whole body-specific IgE antibodies, in more than half of our asthmatic patients. These results showed that it was necessary, further, to study the cross-reactivities between these immune-reactive bands of the insects at molecular basis.