ABSTRACT
Transgenic mice containing intact copies of the human immunodeficiency virus (HIV) proviral DNA were constructed. Founder animals were not viremic for HIV and remained healthy during a 9-month observation period. After being mated with nontransgenic animals, one founder mouse (No. 13) gave rise to F1 progeny that developed a disease syndrome characterized by marked epidermal hyperplasia, lymphadenopathy, splenomegaly, pulmonary lymphoid infiltrates, growth retardation, and death by day 25 of life. Infectious HIV, indistinguishable from parental virus by immunoblot analysis, was recovered from the spleen, lymph nodes, and skin of five of five affected animals.
Subject(s)
Acquired Immunodeficiency Syndrome/microbiology , DNA, Viral , Disease Models, Animal , HIV/genetics , Acquired Immunodeficiency Syndrome/immunology , Acquired Immunodeficiency Syndrome/pathology , Animals , DNA Probes , DNA, Viral/analysis , Epidermis/pathology , HIV/immunology , HIV/isolation & purification , HIV Antibodies/analysis , Lung/pathology , Lymph Nodes/microbiology , Lymph Nodes/pathology , Mice , Mice, Transgenic , Nucleic Acid Hybridization , Skin/microbiology , Skin/pathology , Spleen/microbiology , Spleen/pathologyABSTRACT
Twelve transgenic founder animals retaining intact copies of the infectious molecular clone of human immunodeficiency virus (HIV)-1 were obtained. All the founders appeared healthy during a 9- to 12-month observation period. However, transgenic offspring of one of the founders (female #13), died within the 1st month of life while manifesting several symptoms characteristic of human AIDS. To discover why only one transgenic lineage was affected and why the founder animal in the affected lineage remained healthy while all of her transgenic offspring were diseased, we compared the organization of the transgene in the transgenic lineages. Restriction enzyme analysis showed that the founder no. 13 was a mosaic carrying in each transgenic cell four tandemly arranged copies of the infectious molecular clone. All the units of the tandem repeat appeared to be correctly preserved with the exception of the 3'-most copy, which terminated near the start of human sequences that flank the 3' long terminal repeat (LTR). The unaffected founders and their transgenic litter usually carried a high number of copies of the provirus. The 5' terminus of the transgene in the unaffected animals appeared to be deleted or rearranged. None of the 12 transgenic founders carried a single copy of integrated provirus. We conclude that infectious molecular clone of HIV-1 can be expressed in transgenic mice, and that the mode of proviral integration similar to that seen during the retroviral infectious cycle (i.e., a single-copy provirus) may be incompatible with the postnatal survival.(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Genes, Viral , HIV-1/genetics , Proviruses/genetics , Animals , Cloning, Molecular , Female , Gene Expression , Male , Mice , Mice, TransgenicABSTRACT
A single intraperitoneal injection of 10(-6.5) mol lanthanum chloride/g body wt (44 mg metal/kg body wt) into pregnant mice reduced the number of successful pregnancies and the average litter size. The most susceptible periods of pregnancy were peri-implantation (days 4 and 6) and near-term period (days 14 and 16). Injection of lanthanum during the peri-implantation period resulted in a cessation of pregnancy in 24-43% of females, and injection during near-term period produced the cessation of pregnancy in 36-46% of the females. The average litter size after injection of lanthanum during peri-implantation or near-term periods was reduced to about 75% of the average litter size in the control animals. No external malformations were observed among fetuses. Paradoxically, the exposure of 1-cell stage embryos to 10(-3.0) M and 10(-3.5) M lanthanum chloride in vitro resulted in a significant improvement of the proportion of embryos developing into blastocysts.
Subject(s)
Blastocyst/drug effects , Lanthanum/toxicity , Pregnancy, Animal/drug effects , Animals , Embryo Implantation/drug effects , Embryonic and Fetal Development/drug effects , Female , Fetal Death/chemically induced , In Vitro Techniques , Litter Size/drug effects , Mice , Mice, Inbred ICR , PregnancyABSTRACT
Sixty-nine human oocytes were subjected to various parthenogenetic stimuli but no activation was observed. Analysis of Feulgen-stained DNA (F-DNA) distribution and content showed anomalies of chromosomal material in 36% of oocytes at 24-75 h after retrieval. A significant loss of F-DNA was noticed in apparently normal metaphase II oocytes remaining in culture for 2-3 days.
Subject(s)
DNA/analysis , Oocytes/physiology , Parthenogenesis , Cells, Cultured , Cold Temperature , Ethanol/pharmacology , Female , Fertilization in Vitro , Humans , Hyaluronoglucosaminidase/pharmacology , Male , Meiosis , Oocytes/analysis , Spermatozoa/physiologyABSTRACT
A construct containing the gene for glycoprotein D of herpes simplex virus (HSV-gD), under the control of the simian virus 40 early promoter, was microinjected into single-cell embryos, and four transgenic mouse lines were established. Three were homozygous (lines 75, 111, and 113) and one was hemizygous (line 108) for the HSV-gD gene. Examination of sera revealed that only one of the lines (line 75) spontaneously produced antibody to HSV-gD. Immunization of the other three lines with vaccinia virus-HSV-gD showed that one of them (line 113) responded by making antibody to HSV-gD, whereas the other two (lines 108 and 111) appeared to be immunologically tolerant. Evidence that tolerance was not absolute was obtained by immunization with infectious HSV, which resulted in an antibody response to HSV-gD in some of the animals from line 111. Examination of organs for HSV-gD mRNA revealed transcripts in the tolerant line (line 108) and in the partially tolerant line (line 111), but not in the nontolerant line (line 113), suggesting that the development of immunological tolerance requires active expression of the HSV-gD gene.