单克隆抗M、抗N抗体的研制及初步应用

用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10～15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性和高效价抗 M 和抗 N 抗体的细胞株,并建立了 GM_4H3、GM_4H_4、N_2A_3和 N_2D_(10)细胞株。这些细胞株可持续分泌免疫球蛋白 G 类抗 M、抗 N 抗体。应用这些抗体,通过血凝法、解离法和 ELISA 斑点法,可进行血及血痕的 MN 分型。在血型检验中,优于多克隆抗 M、抗N 血清。     

唾液中H及H2血型物质的定量研究

用抗H1E3（H1及H2特异性）及抗H224（H2特异性）抗体，以时间决定性荧光免疫测定法对129例个体唾液中的H及H2血型物质进行定量检测。证明在分泌型唾液中含有大量H物质，其主体是H1物质，H2物质的含量为H物质的1／10～1／20。在非分泌型唾液中检出少量H1物而无H2物质，表明H2物质的检出与否可作为区分分泌型与非分泌型的标准，推测唾液中H物质含量的差别，由Se．H基因编码的两种α（1→2）岩藻糖基转移酶活性差异所决定。     

人类A、B抗原单克隆抗体的研制和初步应用

作者用人类A、B型红细胞及A、B型分泌型唾液免疫8周龄的BALB/C小鼠,用经免疫的脾细胞与SP2/O骨髓瘤细胞融合,用血凝法筛选出分泌抗A抗体和抗B抗体的细胞株各3株。经一年多的传代,仍然能稳定地分泌抗A、抗B抗体,抗体特异性专一,培养上清液效价能达到2048倍,并初步应用于血液及体液(斑)的检验。     

抗人N单克隆抗体的制备及其特性的研究

用ON型人红细胞免疫BALB/c小鼠脾细胞与SP2/O骨髓瘤细胞融合,获得分泌IgM类抗人N单克隆抗体杂交瘤细胞株(ON_1D_3)。其培养上清液抗体效价为128～256,诱生小鼠腹水的抗体效价为8192;与Panel A红细胞反应呈抗N特异性;检测273名成人红细胞,只特异性凝集N(?)MN型人红细胞,不凝集M型红细胞。与猴等9种动物红细胞无交叉反应;酶修饰试验结果表明,ON_1D_3识别的抗原决定簇是唾液酸依赖性的。     

非分泌型的基因型与血型物质的分泌研究

阐明非分泌型的基因型与血型物质分泌量和Lewis表型的关系。应用时间决定性荧光免疫测定法（TR－FIA），检测传统的A型Lewis阳性非分泌型个体唾液中H、A及Lewis抗原含量，并以序列特异性PCR，确定其基因型。非分泌型个体唾液中检出了高含量的Lea抗原，其中8例不同程度的检出了少量H、A和Leb抗原，其FUTZ位点为se2／se2纯合型，属Le（a＋b＋）型；1例基因型为se3／se5杂合型，未能检出H、A及Leb抗原，属Le（a＋b－）型。se2是弱分泌基因，se3及se5是非分泌基因。     

单克隆抗A、抗B、抗H指示红细胞的研制及应用

<正> 本文报道根据反向间接血凝(RPHA)原理,用单克隆抗A、抗B、抗H抗体分别在合适的pH值醋酸盐缓冲液中,致敏醛化人“O”型红细胞,制得单克隆抗A、抗B、抗H指示红细胞。用来检验人体液斑的ABH血型物质。单克隆抗A、抗B、抗H指示红细胞对相应分泌型人唾液的凝集效价为8192倍、4096倍     

中和法检验人体液斑中H型物质含量在个人识别中的应用4例

中和法检验人体液斑中Ｈ型物质含量在个人识别中的应用４例刘仙洲（河北省顺平县公安局；顺平０７２２５０）Ａ、Ｂ、Ｈ类血型物质不仅存在于红细胞上，在唾液、精斑、阴道分泌液及人体各组织也广泛分布。分泌型人的体液中除分泌与本身相同的型物质外，还可分泌Ｈ型物质，...     

人类分泌状态的研究及进展

ABO血型系统属人类的重要遗传标记之一。ABO抗原不仅存在于红细胞膜，也广泛存在于人体的体液和分泌液中。在体液及分泌液中分泌ABH血型物质的人称作分泌型（Seers－tors，See）。A型人分泌A物质，B型人分泌B物质，O型人分泌H物质。所有分泌型人都有H物质的分泌；不分泌ABH血型物质的人称作非分泌型（nonsecretors，uSe）[1]。分泌ABH血型物质，首先是在检测人精液和唾液时被证实[2]。后来的研究结果表明：在人类分泌型个体的消化道（唾液、胃粘膜、胆汁、胎粪）、泌尿生殖道（精液、阴道分泌物、卵巢囊肿液、尿）、呼吸道及乳汁…     

用酶标抗体免疫组化法测定性交后阴道内容物中精子的ABO血型

应用间接酶标抗体免疫组化法测出了53例新鲜精液、5例陈旧精斑及40例阴道分泌液中的精子与阴道脱落上皮细胞的ABO血型,30例精子与不同血型分泌型阴道分泌液孵育,未发现精子吸附阴道液中血型物质的现象,同时发现人类睾丸曲精细管中部份生精细胞、精子细胞,精子;直细精管部份上皮细胞、精液、精子;睾丸网大部份上皮细胞及副睾管中的精液与精子均含ABH抗原,故认为精子上的ABH抗原主要是精子固有抗原,13例性交后阴道内容物中精子的ABO型测定结果:7例与供者血型吻合,6例不吻合。6例中5例从O型精子中测出了女方分泌型阴道分泌液中的A或AB物质,1例B型精子未测出B及H抗原,文中对这种现象进行了讨论。     

免疫抗A、抗B、抗H沉淀素血清的制备

本文介绍了用浓缩的分泌型人唾液免疫鸡,经吸收精制获得鸡抗A、抗B、抗H沉淀素血清的方法。用本法制备的抗血清,对相应血型人唾液的特异性沉淀价可达1:512倍以上,对非相应血型者的分泌液不出现非特异性沉淀反应。应用该血清进行环状沉淀试验、单向琼脂扩散试验等免疫学试验,可简便、准确地鉴定人唾液斑、精液斑等分泌液斑的ABO血型。     

Immunoenzyme determination of ABO and secretor status in paraffin-embedded autopsy material

Using the indirect immunoperoxidase technique (PAP method), A, B, and H antigens were identified on formaldehyde-fixed, paraffin-embedded kidney tissue from 100 autopsies. Comparison with the serologic findings showed all our blood group determinations to be correct. The labeling of the collecting tubules was evaluated as characteristic of the secretor. The secretor status determined according to this parameter was unequivocally confirmed by the Lewis constellation in 78 of 82 cases; group Le(a-b-) could be differentiated with immunohistochemical methods in secretors and nonsecretors. Determination of ABO blood groups and secretion behavior with immunohistochemical methods was correct even in those cases where classic serology failed due to hemolysis and decomposition. Immunohistologic results obtained with monoclonal antibodies were better than those obtained with human sera.     

Determination of ABH secretor status by an electronic quantitation method

Blood Group A and B substances in secretor (Se) and nonsecretor (se) salivas were tested by means of an electronic data processing-hemagglutinin-inhibition test (EDP-HAIT) with immunoglobulin M (IgM) isohemagglutinins. Besides a difference in quantity, the blood group substances in Se saliva showed high binding efficiencies compared with those in se saliva. EDP-HAIT with IgG isohemagglutinins proved no difference in the binding efficiencies of Se and se salivas. The determination of secretor status by EDP-HAIT with IgM isohemagglutinin was accurate because the conclusion was obtained based on two different quantitative results. Secretor status of some salivas in gargled water could be determined by comparing the binding efficiencies.     

Immunohistological studies on ABH-activities in secretory cells of human major salivary glands--correlation between ABH-activities in the secretory cells and secretor-nonsecretor

The activities of A, B and H in serous cells (S-cells), mucous cells (M-cells) and excretory duct cells were examined in a large number of paraffin sections of three major salivary glands obtained from 91 corpses, using the immunofluorescence technique. The results are: By taking H activity in S-cells of the submandibular gland or A, B and H activity in M-cells of the sublingual gland as an indicator, the salivary glands were classified as Type I showing activity and Type II showing no activity. No glands corresponding to the intermediate type, as seen in the case of saliva, were noted at all. Among 91 corpses, 70 cases were classified as Type I and 21 as Type II. The results matched well with those of Lewis type tested on blood. The frequencies of the typing (Type I; 76.9%, Type II; 23.1%) were approximately in concordance with those of secretor and nonsecretor in Japanese saliva. From these results, it was assessed that the former corresponded to the secretor type in the case of saliva, and the latter to the nonsecretor type. Even in the same individual, both S-cells and M-cells exhibited different productivities of substances, depending on the glands to which they belonged. Namely, only S-cells in the submandibular gland belonging to Type I showed only H activity independent of the blood group of the individual, but the other S-cells in the other major glands did not show any activity for A, B and H. M-cells exhibited strong activity for H and/or A and/or B in the sublingual and submandibular gland and belonged to Type I, but little activity in the sublingual gland belonged to Type II. In the submandibular gland of Type II, some M-cells showed activity and others did not. On the basis of the above results, we discuss the applicability of the present genetic theory concerning the secretor and nonsecretor type in saliva to salivary glands and cells, and further refer to the reasons for appearance of the weak secretor type or intermediate type in saliva.     

唾液血型抗原分泌时限变化及温度对抗原活性的影响

探讨人唾液中ABH血型抗原不同时限的分泌量,以及保存温度对血型抗原活性的影响。应用时间决定性荧光免疫测定法（TR．FIA）对O型分泌型10例和非分泌型5例人在不同条件下唾液中H抗原量进行检测。唾液血型抗原的分泌量随时间而波动,进餐后降低明显,但不干扰分泌型的判定。37℃保存48h抗原活性完全丧失,6℃保存1周抗原活性几乎没有变化。结果表明,唾液分泌时段不影响分泌型判定,将唾液制成斑痕可长期保存样品。     

ABO typing studies on liquid urines

ABO typing was successfully performed on 46 urine samples whose ABO group and secretor status had been determined previously from blood and saliva. Twenty-four urine samples were collected on which blind studies, time studies, and storage studies were performed. Multiple urines from several individuals were collected to evaluate the duplicity of the test. Also, urines were collected from pregnant and menstruating females to determine if ABO typing was affected under these conditions. Results of these studies are discussed.     

The rapid determination of the ABO group from body fluids (or stains) by dot enzyme-linked immunosorbent assay (dot-ELISA) using enzyme-labeled monoclonal antibodies

Using ABH enzyme-labeled monoclonal antibodies, the authors could rapidly detect the ABO group from body fluids and body fluid stains by the dot enzyme-linked immunosorbent assay (dot-ELISA). In this test, the antigen was immobilized on nitrocellulose paper; the entire piece of paper was coated with an appropriate dilution of enzyme-labeled McAb directly against the antigen of interest; and, finally, 3,3'-diaminobenzidine (DAB) substrate solution was added. The site of a positive reaction is clearly visible as a brown spot. We analyzed 521 samples and got satisfactory results. We also analyzed 99 practical case samples by this method and achieved the same results as those obtained by other researchers using other methods. This method is accurate, simple, direct, rapid, and sensitive; it also produces easily observed results, requires no equipment, and can be completed in 30 min. The test proved to be clearly more sensitive for the detection of the ABO blood group in secretor saliva than the conventional hemagglutination inhibition test. Also saliva diluted 10(-4) to 10(-5) and the ABO group of nonsecretor saliva and urine could be easily detected by this method.     

The ABH reactions of seminal stains

The ABO grouping results from approx. 1000 seminal stains have been collected and analysed. Most of the stains came from rape cases where the ABO and secretor status of both complainant and suspect were known. The results of the survey provided information concerning the usefulness of elution and inhibition as methods of body fluid grouping, the relative strengths of reaction of the A, B and H antigens in body fluids and the interpretation of the ABH reactions of body fluid stains.     

A, B and H group specific substances in human vitreous humor

The presence of A, B and H group specific substances in vitreous humor taken from 105 human corpses was determined. Good agreement was obtained between these group substances and the ABO blood group. The relationship with the secretor type is discussed.     

H-抗原缺乏分泌型个体FUT1及FUT2等位基因的分子基础

一例H-缺乏分泌型个体被发现。其血清学表现为：红细胞上无A、B、H抗原；唾液中有B、H抗原分泌；血清中检出了抗A抗体。推测其为类孟买OHmB型。在该个体FUT1等位基因编码区上发现两处单碱基突变（T460C、G1042A）。这两个点变异，将导致两个氨基酸的置换（Y154H、E348K）。同时也破坏了限制性内切酶RsaⅠ和AvaⅠ的作用部位。用PCR－RFLP法可检出此两种变异。用PCR－RFLP法证实，该个体为T460C、G1042A变异的纯合子。在136例随机个体中未能查出上述变异。将该FUT1基因转染COS－7细胞未能检出α2－FUT活性及证实H抗原的表达。该个体的FUT2基因与野生型一致。