Gene therapy of malignant brain tumors: a rat glioma line bearing the herpes simplex virus type 1-thymidine kinase gene and wild type retrovirus kills other tumor cells.

Tumor cells infected with a retrovirus vector (VIK) containing the herpes simplex virus thymidine kinase (HSV-TK) gene can be selectively killed by treatment with nucleoside analogues, such as ganciclovir. To mediate delivery of the HSV-TK gene to "recipient" tumor cells, "donor" C6 rat glioma cells infected with the VIK vector (C6VIK) were superinfected with wild type Moloney murine leukemia virus (WT Mo-MLV). These modified donor cells (C6VIKWT) produced both wild type retrovirus and the VIK vector. In culture, C6VIKWT cells were 300-fold more sensitive to the toxicity of ganciclovir than were C6VIK cells, suggesting that the presence of wild type retrovirus contributed to the toxicity. Co-culture of C6VIKWT cells with the C6 subline, C6BAG, sensitized the latter to ganciclovir treatment. Nude mice inoculated subcutaneously with a mixture of C6VIKWT and C6BAG cells showed regression of subsequent tumors when treated with ganciclovir. The observations show that tumor cells modified in culture by infection with a retrovirus bearing the HSV-TK gene and wild type retrovirus are not only sensitive to ganciclovir, but can transfer this sensitivity to neighboring "naive" tumor cells in culture and in vivo.     

Restriction endonuclease analysis of varicella-zoster vaccine virus and wild-type DNAs

The DNA from several clinical isolates of varicella-zoster virus (VZV) were compared with the DNA from the vaccine strain VZV using three restriction endonucleases: BamHI, BgII, and HpaI. When electrophoresed through an agarose gel, the vaccine DNA digestion pattern was significantly different from the digestion patterns of the wild-type DNAs. Variations in the digestion pattern of the separate clinical isolates were also observed.     

脂质体转染AFP基因构建树突状细胞肝癌瘤苗及体外活性检测

目的：探讨以ＡＦＰ为靶点，构建ＡＦＰ－ＤＣ肝癌瘤苗及其抗肝癌免疫治疗的可能性。方法：应用脂质体将ＡＦＰ基因转染体外培养的未成熟 ＢＭＤＣ，构建 ＡＦＰ－ＤＣ肝癌瘤苗，以流式细胞术、Ｗｅｓｔｅｒｎ ｂｌｏｔ、３Ｈ－ＴｄＲ法和 ＭＴＴ法等检测其免疫活性。结果：ＡＦＰ－ＤＣ瘤苗不仅能产生和分泌ＡＦＰ，而且能上调自身的Ｂ７分子和ＭＨＣ分子，明显刺激Ｔ细胞增殖及提高ＣＴＬ的杀伤作用。结论：提示肝癌相关基因ＡＦＰ可作为抗肝癌基因治疗的切入点。     

Sequence Analysis of the Washington/1964 Strain of Human Parainfluenza Virus Type 1 (HPIV1) and Recovery and Characterization of Wild-Type Recombinant HPIV1 Produced by Reverse Genetics

A complete consensus sequence was determined for the genomic RNA of human parainfluenza virus type 1 (HPIV1) strain Washington/20993/1964 (HPIV1 WASH/64), a clinical isolate that previously was shown to be virulent in adults. The sequence exhibited a high degree of relatedness to both Sendai virus, a PIV1 virus recovered from mice, and human PIV3 (HPIV3) with regard to cis-acting regulatory regions and protein-coding sequences. This consensus sequence was used to generate a full-length antigenomic cDNA and to recover a recombinant wild-type HPIV1 (rHPIV1). Interestingly, the rHPIV1 could be rescued from full-length antigenomic rHPIV1 cDNA using HPIV3 support plasmids, HPIV1 support plasmids, or a mixture thereof. The replication of rHPIV1 in vitro and in the respiratory tract of hamsters was similar to that of its biologically derived parent virus. The similar biological properties of rHPIV1 and HPIV1 WASH/64 in vitro and in vivo, together with the previous demonstration of the virulence of this specific isolate in humans, authenticates the rHPIV1 sequence as that of a wild-type virus. This rHPIV1 can now be used to study the biological properties of HPIV1 and as a substrate to introduce attenuating mutations for the generation of live-attenuated HPIV1 vaccine candidates.An erratum to this article can be found at     

IL—6基因转染的瘤苗体内诱导的抗肿瘤免疫功能与效果

经丝裂霉素C体外处理后,将IL-6基因转染的、高分泌的IL-6的B16黑色素瘤细胞制成瘤苗。结果发现,体内注射IL-6基因转染的瘤苗后,小鼠脾脏CTL活性、NK活性及IL-2诱导的LAK活性显著升高。经IL-6基因转染瘤苗体内治疗后,荷瘤小鼠的皮下肿瘤生长显著减慢、肺转移结节数显著降低、存活期显著延长,若同时合用低剂量IL-2,则上述治疗效果更好。可见IL-6基因转染的瘤苗能有效地通过诱导机体抗肿瘤免疫功能而发挥抗肿瘤作用,与低剂量IL-2合用后,IL-6基因转染的瘤苗的抗肿瘤效果更佳。     

The immune response of healthy adults to a reduced dose of hepatitis B vaccine

Three hundred thirty-six medical personnel from hemodialysis centers were treated with three doses, 20 μg each, of the Merck hepatitis B vaccine (at 0, 1, and 6 months). Within 1 month after the first injection, 41% converted to anti-HBs positivity; after the second injection the conversion rate rose to 80–90%; and after the booster, to 96–98%. The later rate remained unchanged during the 18-month follow-up period. Only 2.8% of those vaccinated did not respond to the vaccine. The patterns of immune responses to 20-μg doses were found to be exactly the same as to 40-μg doses. It is suggested that reduced doses of vaccine should be as efficacious as the larger ones.     

Immunization with Combined HSV-2 Glycoproteins B2:D2 Gene DNAs: Protection against Lethal Intravaginal Challenges in Mice

The immunity of a combined DNA vaccine of HSV-2 glycoproteins B2 (gB2) and D2 (gD2) genes in comparison to individual vaccines was studied with regard to protecting against the HSV infection. Two recombinant DNA vaccines of the pHS2-gB2 or pHS2-gD2 were constructed and formulated. The neutralizing antibody titers appeared higher in the B2:D2 gene cocktail-vaccinated mice than that of the individual B2 or D2 gene-vaccinated group alone, and the positive KOS control induced higher titer of the neutralizing antibody than combined or individual gene vaccines. The mock-immunized mice failed to induce enough. The ranks for the CTL activity and the protection rates against the lethal intravaginal challenge were shown as KOS>B2:D2 cocktail>D2>B2 gene vaccines. The vaginal external diseases in the B2:D2 or D-vaccinated mice were significantly reduced against the challenging dosages. The virus titers in the vaginal secretions of the vaccinated mice significantly reduced with time, and the B2:D2 gene vaccine decreased more than each individual vaccine alone. It can be concluded that the cocktailed vaccines are more effective in the humoral and cellular immune responses in the mice, and in the protection of the mice against the intravaginal challenging dosages when compared with individual gene vaccines. All the DNA vaccines failed to block the latent infection in sensory nerves.     

大鼠抗—HBs的单克隆抗独特型抗体杂交瘤细胞系的建立

用纯化的小鼠单克隆抗—HBs和纯化的大鼠多克隆抗—HBs免疫LOU/C大鼠,取免疫大鼠脾细胞与IR983F骨髓瘤细胞融合,获得两株(TD_1、TD_2)持续分泌抗—HBs的单克隆抗独特型抗体(抗—Id,Ab_2)杂交瘤细胞系。对TD_2进行了较全面的鉴定,TD_2的Ig类型为IgG,亚类为IgG_(2?),核型分析结果染色体数为65条。通过ELISA竞争抑制和中和抑制试验表明,TD_2所分泌的Ab_2既能被不同来源的抗—HBs所中和。又能被HBsAg.所竞争,且都呈现剂量依赖关系。将纯化的TD_2IgG免疫同系大鼠,诱导出了具有抗—HBs活性的抗—抗—独特型抗体(Ab_3)。上述结果证实,TD_2所分泌的单克隆抗体(McAb)是带有HBsAg表位内影像,具有HBsAg免疫源性的抗—HBs的单克隆抗—Id。     

Histopathological studies on the local reactions induced by complete Freund's adjuvant (CFA), bacterial lipopolysaccharide (LPS), and synthetic lipopeptide (P3C) conjugates

The inflammatory reactions following subcutaneous application of adjuvants revealed characteristic pathological patterns. The injection of complete Freund's adjuvant (CFA) resulted in the formation of large lipid deposits encircled by an inflammatory reaction and concentrically arranged collagen bundles. Bacterial lipopolysaccharide (LPS) caused granulomatous aggregations of mononuclear cells with thrombotic vessel occlusions. Inoculation of the lipopeptide adjuvants induced accumulation of mononuclear cells with only minimal fibrotic changes which were resolved after day 28. Lipopeptide conjugates based on the head group tripalmitoyl-S-glyceryl-cysteinyl-serin (P3CS) can thus be used as effective immunogens and adjuvants without long-term tissue damage.     

大鼠保留交感神经的高选择性迷走神经切断术实验模型的建立

目的：建立保留交感神经的高选择性迷走神经切断术（ＨＳＶ－ＡＰ）的动物实验模型。方法：６０只雄性ＳＤ大鼠随机分为三组：ＨＳＶ－ＡＰ组、ＨＳＶ组和正常对照组（Ｃ组），每组２０只。结果：ＨＳＶ－ＡＰ术后胃壁泌酸区交感神经分布密度与Ｃ组比较，下降不显著，去甲肾上腺素含量与Ｃ组无差别（７４５．０±４０８．９ＶＳ８９９．６±２３５．１，Ｐ＞０．０５）。而ＨＳＶ术后胃部泌酸区交感神经分布密度显著下降，去甲肾上腺素含量低于ＨＳＶ－ＡＰ术后（２９３．０±２１４．８ＶＳ７４５．８±４０８．９，Ｐ＜０．０１）。ＨＳＶ－ＡＰ术后胃体部迷走神经追踪脑干内未见标记细胞，而胃窦部迷走神经追踪脑干内可见标记细胞。结论：本试验证实采用保留胃小弯血管的ＨＳＶ—ＨＳＶ－ＡＰ术式，既可完整切断支配胃部泌酸区的迷走神经，又可保留交感神经。