体外免疫法抗MG7人源性单抗的研制及其意义

本文建立了人淋巴细胞体外致敏技术,用本文作者既往制备的鼠抗人胃癌单抗MG7作为免疫原,在体外致敏人淋巴细胞获得成功。经与本文作者在建立的人鼠种间骨髓瘤细胞系FMC-1进行人鼠人双杂交,获得一株能与鼠源性抗体反应,但不与人、羊、马、兔等其他种属动物产生的抗体相反应的人源性单抗HMG7。本文讨论了HMG7可能的应用价值,以及在人淋巴细胞体外致敏过程中应注意的几个环节。     

人-鼠杂交瘤制备人单克隆抗体技术的研究

本文采用小鼠骨髓瘤细胞系(Sp2/0)与破伤风类毒素(TT)免疫的人外周血淋巴细胞(PBL)进行融合,对人—鼠杂交瘤细胞系制备人单克隆抗体的技术条件进行了探讨。按免疫时间(d)及体外刺激与否分为4组。结果表明:(1)PWM+TT体外刺激可导致B细胞总数增加,且融合率高,容易建成稳定分泌人单抗的细胞株。(2)经TT免疫的人PBL,6d就有抗TT抗体(IgG)的前体细胞出现。15d及21d时,抗TT抗体前体细胞的频率更高。(3)体内免疫7d及14d并分别经体外刺激的两组共建立5株阳性株。经双扩碓定IgG2株,IgM3株。冻存10个月后复苏、传代及适当的克隆化后仍为阳性。上清液中人Ig的分泌量为0.42～1.15μg/ml。经染色体核型鉴定,碓认为人-鼠杂种细胞。     

体外免疫法抗MG7人源性单抗的研制及意义

人源性单抗用于人体疾病的体内诊断及治疗,不仅免疫原性弱,半衰期长,且能介导某些重要的免疫反应,以调节机体的免疫衡稳状态或增加抗体的抗病能力。但是,由于许多抗原物质如肿瘤抗原不能直接注入人体进行免疫,故给人单抗的制备带来困难。本文在国内首次建立人淋巴细胞体外致敏技术,并用本文作者既往制备的鼠抗人胃癌单克隆抗体MG7作为免疫原,在体外致敏人淋巴细胞获得成功。经与本文作者在国内首次建立的人鼠种间骨髓瘤细胞系FMC-1进     

人-鼠杂交瘤中染色体与单抗泌稳定性的关系

人-鼠杂交瘤产生人单克隆抗体是一项很有应用前景的技术,它是由有基因缺陷的小鼠骨髓瘤细胞和人的B 淋巴细胞融合,所得的杂交瘤可以在体外长期产生有着抗原特异性的人单抗。但是这种异种杂交瘤却十分不稳定,在杂交瘤形成后的很短时间内即丢掉人抗体的分泌能力。有人认为这是由于人染色体的丢失,是由于调节基因的失活,也有人认为这和融和所用的亲本细胞系有关,本文综述了染色体与抗体分泌稳定性的关系以及与染色体分析相关的技术。     

人鼠种间骨髓瘤细胞系FMC-1的建立及在人单抗制备中的应用

本文报告在国内首次建立的一株能用于制备人源性单抗的人鼠种间骨髓瘤细胞系FMC-1。FMC-1对8-偶氮鸟嘌呤及鸟本苷具有抗药性,最大耐受剂量分别可达64ng/ml和1×10~(-4)     

用免疫荧光法筛选抗人μ链单克隆抗体(McAb)的研究

用人血清IgM球蛋白重链-μ链免疫的BALB/c小鼠脾细胞与鼠骨髓瘤Sp 2/0细胞在PEG作用下融合,用免疫荧光法筛选出6种分泌McAb的杂交瘤细胞,其中5株确定为抗人μ链McAb,一株未定。杂交瘤细胞经四次克隆,半年多传代,接种BALB/c鼠可稳定的产生腹水抗体。     

4株鼠抗人B7-1分子单抗的制备及其生物学活性的鉴定

目的：制备鼠抗人B7—1分子(mAb)单抗及研究其生物学活性。方法：以人多发性骨髓瘤细胞转人B7-1基因细胞株XG7-B7为免疫原，采用B淋巴细胞杂交瘤技术制备鼠抗人B7—1分子单抗；以Western blot及间接免疫荧光标记法鉴定其特异性和亲和力；采用^3H—TdR掺入实验和Annexin-Ⅴ染色分析测定该单抗的生物学功能。结果：获得了4株持续分泌抗人B7—1分子的特异性单抗的杂交瘤细胞株，分别命名为1F11、3H8、6H2和7B10，其分泌的单抗类别分属于小鼠IgG1和IgM；4株单抗均具有良好的特异性和亲和力；1F11、3H8和6H2能部分组断B7—1分子基因转导细胞介导的共刺激信号，从而抑制T细胞的增殖效应(抑制率21％—52％)；且能诱导天然表达B7—1分子的人B系淋巴瘤细胞Raji的凋亡。结论：成功研制4株功能性抗人B7—1分子抗体，这些抗体在抗异体组织器官移植排斥反应及在B系淋巴瘤的治疗中具有潜在的应用价值。     

体外致敏制备人单克隆抗体研究现状

体外致敏人淋巴细胞,诱生大量抗原特异性B 细胞和骨髓瘤细胞融合,这是人单抗制备研究中的一项关键步骤。体外致敏提高了杂交瘤形成率和抗体分泌稳定性,推动了人单抗研究的进展。成功的体外致敏有赖于淋巴细胞来源、细胞组成、抗原性质和剂量、血清、分裂原、生长因子、佐剂、免疫时程等因素。本文就近年来这方面的研究现状作一介绍。     

EBV-杂交瘤技术制备抗乳癌人单抗的研究

本研究用EBV-杂交瘤技术制备抗乳癌人单抗获得初步成功。实验中首先预选出病人血清对肿瘤相关抗原有强阳性反应的乳癌患者,取其引流区淋巴结细胞,去除T细胞。在体外用EBV转化。筛选出分泌阳性抗体的B细胞,扩增后与一株人-鼠骨髓瘤杂交株HMD-33融合,用含HAT-ouban 1640培基进行选择培养。融合后两周左右见有杂交瘤生长,融合率为38％,从中获得16株稳定分泌抗体的杂交瘤,抗体类型75％为人IgM余为人IgG或混合型,上清     

抗人T细胞白血病1型病毒(HTLV1)的人源单克隆抗体

本文报告了用EBV-杂交瘤技术产生人单克隆抗体。从多发性硬化症病人外周血获得B淋巴细胞,经EB病毒转化,建立了永久性的B细胞系并与小鼠骨髓瘤细胞(P_3×63Ag8.653)或Spatz-4(人与鼠细胞杂交的瘤细胞系)融合,用ELISA方法筛选阳性杂交瘤,再经亚克隆培养,筛选出抗HTLV1特异性抗体,通过SDS-PAGE,western blot及FACS分析,证明这些抗体只与HTLV1的24kDa蛋白反应,并全部分泌IgM类型免疫球蛋白。     

Analysis of pairs of individual Ia-E.P.S.P.S in single motoneurones.

1. Recordings of individual e.p.s.p.s evoked by the action of single medial gastrocnemius Ia fibres have been made from medial gastrocnemius motoneurones. In many motoneurones the action of two Ia fibres has been observed and the properties of the e.p.s.p.s compared. 2. For sixty-three pairs of averaged e.p.s.p.s, each from the same motoneurone, the ratio of half-widths was plotted against the ratio of rise times. These results were compared with theoretical values derived from the Rall compartmental model. It was found that variations in synaptic current time courses and differences in the termination of localized synaptic terminals were not sufficient to account for all the data. 3. Amplitude and rise time were inversely related but the correlation coefficient was very low. For pairs of e.p.s.p.s in the same motoneurone the e.p.s.p. with the fast rise time was larger than that with the slow rise time in forty-eight of sixty-three cases. 4. In a given motoneurone individual e.p.s.p.s evoked by the action of different Ia fibres did not vary greatly in amplitude. The ratio of peak amplitudes was less than 3 for 86% of the pairs of e.p.s.p.s examined, and the maximum was 4-8. 5. Amplitude histograms were constructed for individual e.p.s.p.s at thirty-three synapses. Twenty-two of them could be shown to satisfy the Poisson law. The others satisfied the binomial law or neither. 6. Within a given motoneurone the amplitude of an e.p.s.p. is closely related to the mean number of quanta released but not to the amplitude of the unit e.p.s.p. produced by the action of a single quantum of transmitter.     

Prevention of E.N.T. diseases

World Health Organization has announced 'HEALTH FOR ALL BY YEAR 2000'. Unfortunately the emphasis at present is only on early diagnosis of a disease and its treatment. The modern investigations and treatment has become very costly which a poor man cannot afford. Preventive medicine is less costly. The developing countries have many socio-economic problems of population explosion, poverty, ignorance, illiteracy, shifting of population from rural to urban areas for employment and traffic jams in cities. Rapid industrialization has resulted in indiscriminate cutting of trees and forests and increasing pollution. Under the present circumstances one wonders whether it would be possible to achieve the WHO's object of health for all or will it remain a dream! The role of 'Yoga and Meditation' and vegetarian diet has been recognised world over for maintenance of normal physical and mental health; these help to lessen mental tension in the present competitive world full of stress and strain for all of us. 3-4% of people in the world avail the beneficial effects of alternative system of medicine is complete as each has its own merits and limitations. Hence our aim should be to choose an integrated system of medicine which would help in preventing diseases and treat them early by a safe and effective procedure. In the present political situation in our country none of the parties have announced any definite programme for population control, through it is a grave problem. We will not be able to fulfil our object of reaching our target of Health for all without strict population control major. Unless this programme is undertaken seriously as a priority in a co-ordinated disciplined manner by all of us and completed in a given period of time, the younger generation will not forgive us. We hope that as true citizens of India we will not spare any efforts on our part in attaining this objective for a bright future.