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Biomedical subjects

R N Lausch

Publications and source records attributed to R N Lausch.

At least 19 recordsLinked to original sources
PubMed1991

Anti-glycoprotein D monoclonal antibody protects against herpes simplex virus type 1-induced diseases in mice functionally depleted of selected T-cell subsets or asialo GM1+ cells.

Passive transfer of a monoclonal antibody (MAb) specific for glycoprotein D (gD) is highly effective in preventing the development of herpes simplex virus type 1-induced stromal keratitis. In the present study, we investigated whether animals which had been functionally depleted of T-cell subsets or asialo GM1+ cells would continue to be responsive to MAb therapy. BALB/c mice were depleted of CD4+, CD8+, or asialo GM1+ cells by treatment with anti-L3T4, anti-Lyt 2.2, or anti-asialo GM1 antibodies, respectively. Functional depletion of CD4+ cells was documented by the loss of delayed-type hypersensitivity responsiveness, while CD8+ cell depletion was accompanied by abrogation of cytotoxic lymphocyte activity. Anti-asialo GM1 treatment led to the loss of natural killer cell lytic activity. Mice depleted of the desired cell population and infected on the scarified cornea with herpes simplex virus type 1 uniformly developed necrotizing stromal keratitis by 3 weeks postinfection. A single inoculation of anti-gD MAb (55 micrograms) given intraperitoneally 24 h postinfection strongly protected hosts depleted of CD4+ cells against stromal keratitis. Likewise, antibody treatment in CD8+ or asialo GM1+ cell-depleted hosts was as therapeutically effective as that seen in non-cell-depleted mice. We also observed that in cell-depleted mice, the virus spread into the central nervous system and caused encephalitis. The CD4+ cell-depleted mice were the most severely affected, as 100% developed fatal disease. Anti-gD MAb treatment successfully protected all (32 of 32) CD4+-, CD8+-, or asialo GM1(+)-depleted hosts against encephalitis. We therefore conclude that antibody-mediated prevention of stromal keratitis and encephalitis does not require the obligatory participation of CD4+, CD8+, or asialo GM1+ cells. However, when mice were simultaneously depleted of both CD4+ and CD8+ T-cell subsets, antibody treatment could not prevent fatal encephalitis. Thus, antibody can compensate for the functional loss of one but not two T-lymphocyte subpopulations.

Animals↗
PubMed1991

Differences in the capacity of two herpes simplex virus isolates to spread from eye to brain map to 1610 base pairs of DNA found in the gene for DNA polymerase.

A intertypic recombinant, designated HSV-R(D1), had previously been generated from non-neuroinvasive HSV-2(186) and neuroinvasive HSV-1(17). Although the recombinant contained less than 2% of the HSV-1 genome, it retained the neuroinvasive phenotype. The nucleotide sequences responsible for the neuroinvasiveness of HSV-R(D1) were previously mapped to a 3.0 kb segment of DNA located within the DNA polymerase gene (mu 0.414 to 0.430) via marker rescue experiments. We have now sequenced this region and compared our results to the published nucleotide sequence of the HSV-1(17) and HSV-2(186) DNA polymerase genes. It was found that the 3.0 kb HSV-R(D1) DNA fragment consisted entirely of HSV-2(186) nucleotide sequences except for the presence of 1610 bp of HSV-1(17) DNA. The 1610 bp of HSV-1 DNA coded for a 536 amino acid (AA) region which were located between AA 254 and 790 of the DNA polymerase enzyme. Comparison of the 536 AA sequence of neuroinvasive HSV-1(17) with the homologous area of the non-neuroinvasive HSV-2(186) DNA polymerase indicated that the two polymerases differed at 56 AA positions. In addition, this area of the HSV-1(17) DNA polymerase was 5 AA acids shorter than the HSV-2(186) DNA polymerase. Specific amino acid changes that might account for the neuroinvasive phenotype of HSV-R(D1) are discussed.

Amino Acid Sequence↗
PubMed1991

Evidence endogenous interferon production contributed to the lack of ocular virulence of an HSV intertypic recombinant.

An intertypic recombinant isolated from rabbit kidney cells following co-transfection of HSV-1(17) and HSV-2(186) DNA failed to induce overt ocular pathology when inoculated onto the murine sacrificed cornea at concentrations as high as 10(7) PFU per eye. In contrast, both parents induced corneal disease at a 1000-fold lower dose. The reason(s) for the failure of the intertypic recombinant, designated RO25X, to induce corneal pathology was investigated. It was found that the recombinant was 100-fold more sensitive to the inhibitory effects of interferon (IFN) alpha/beta than the parent strains in corneal button growth studies in vitro. R025X readily grew in cultured mouse corneal fibroblasts at a low multiplicity of infection. However, the peak titer was approximately 8-fold lower than that of strain 17. Addition of rabbit anti-IFN alpha/beta to the culture medium resulted in a 4 to 5-fold increase in infectious titer compared to its growth in the absence of antiserum. Most significantly, when mice were pre-treated in vivo with anti-IFN alpha/beta 24 hours prior to virus corneal infection, 67% of the recipients developed moderate to severe stromal keratitis, whereas none of the controls developed corneal pathology. Blepharitis was also significantly increased in incidence and severity in the antiserum treated hosts. We conclude that the inability of R025X to induce ocular disease was due, at least in part, to the inhibitory effects of interferon produced in response to infection.

Animals↗
PubMed1990

Nucleotide sequences responsible for the inability of a herpes simplex virus type 2 strain to grow in human lymphocytes are identical to those responsible for its inability to grow in mouse tissues following ocular infection.

A study was undertaken to determine whether genes associated with herpes simplex virus (HSV) neuroinvasiveness in mice influence the growth of HSV in man, the virus's natural host. HSV-2(186), a nonneuroinvasive HSV strain, was found to replicate poorly (less than 3-fold) in cultures of phytohemagglutinin (PHA) stimulated human peripheral blood mononuclear cells (PBMC). In contrast, seven other HSV strains all multiplied 40- to 100-fold. The paucity of HSV-2(186) growth in PBMC was not due to a failure of this strain to grow in primary human cells because high titers (greater than 10(8) PFU/ml) were obtained following infection of human foreskin fibroblasts. The genetic basis for the deficient growth was analyzed by marker rescue experiments. Recombinant HSV-2 strains were generated in marker rescue experiments utilizing HSV-2(186) DNA and plasmids containing a cloned DNA polymerase gene isolated from a neuroinvasive HSV strain possessing the capacity to replicate in human PBMC. Progeny which rescued DNA from the cloned HSV DNA polymerase gene replicated 40- to 100-fold in PHA-stimulated PBMC. Moreover, unlike the HSV-2(186) parent, HSV-2(186) isolates possessing rescued DNA grew well in the eye, trigeminal ganglion, and brain of mice and induced fatal encephalitis. The results indicate that nucleotide sequences responsible for increasing the capacity of HSV-2(186) to grow in PBMC of man are identical to those responsible for increasing the capacity of this strain to grow in mouse tissues and to spread from the eye to the brain.

Adult↗
PubMed1990

Ocular avirulence of a herpes simplex virus type 1 strain is associated with heightened sensitivity to alpha/beta interferon.

BALB/c mice infected on the scarified cornea with herpes simplex virus type 1 strain 35 [HSV-1(35)] rarely developed ocular disease even at challenge doses as high as 10(7) PFU per eye. In contrast, HSV-1(RE) consistently induced stromal keratitis at an inoculum of 2 x 10(4) PFU. The goal of this study was to determine the reason for the difference in virulence between the two HSV strains. Both HSV-1 strains replicated to similar titers in excised corneal "buttons." However, after in vivo infection of the cornea, the growth of strain 35 was evident only during the first 24 h postinfection, whereas the replication of strain RE persisted for at least 4 days. In vitro tests revealed that HSV-1(35) was greater than 10 times more sensitive to alpha/beta interferon (IFN-alpha/beta) than HSV-1(RE). Both strains induced comparable serum levels of IFN after intraperitoneal inoculation. The kinetics of HSV-1(35) clearance from the eye was markedly altered by treatment with rabbit anti-IFN-alpha/beta. Virus titers exceeding 10(4) PFU per eye could be demonstrated 4 to 5 days postinfection in mice given a single inoculation of antiserum 1 h after infection. Furthermore, anti-IFN treatment in 3-week-old mice infected with HSV-1(35) led to the development of clinically apparent corneal disease which subsequently progressed to stromal keratitis in the majority of recipients. These results indicate that the striking difference in the capacity of HSV-1(35) and HSV-1(RE) to induce corneal disease was related to the inherently greater sensitivity of strain 35 to IFN-alpha/beta produced by the host in response to infection.

Animals↗
PubMed1990

Effective antibody therapy in herpes simplex virus ocular infection. Characterization of recipient immune response.

The immunotherapeutic potential of a monoclonal antibody specific for glycoprotein D of herpes simplex virus was evaluated in a murine ocular infection model. Passive transfer of antibody at microgram concentrations was able to promote resolution of corneal opacity and hasten healing of blepharitis. Antibody treatment did not prevent development of either a cellular or humoral antiviral immune response. In fact, kinetic studies revealed that the early delayed-type hypersensitivity response was significantly more vigorous in the treated group than in the controls. Potential explanations as to how a single microgram inoculation of antibody could exert a therapeutic effect are discussed.

Animals↗
PubMed1989

Quantitation of purified monoclonal antibody needed to prevent HSV-1 induced stromal keratitis in mice.

A purified IgG2a monoclonal antibody with a neutralizing titer of 10(4) and specificity for gD was evaluated for its therapeutic potential in a murine ocular infection model. BALB/c mice, infected on the scarified cornea with 10 times the HSV-1 strain RE concentration needed to produce severe and persistent stromal opacity, were given a single inoculation of antibody intraperitoneally 24 hours later. The animals were then followed for corneal disease development. Antibody, at concentrations as low as 10 micrograms per mouse, was strikingly effective at preventing corneal opacity. Furthermore, the corneas, once clear, remained clear whereas the controls developed +3 to +5 stromal disease which was still present 60 days post-infection. Animals that had been treated and recovered from infection were resistant to subsequent HSV-1 challenge on the opposite cornea. These results demonstrate the therapeutic potential of systemically administered microgram quantities of anti-gD antibody.

Animals↗
PubMed1988

Evidence that the gene for herpes simplex virus type 1 DNA polymerase accounts for the capacity of an intertypic recombinant to spread from eye to central nervous system.

HSV-1(17) replicates 100-fold more efficiently than HSV-2(186) within trigeminal ganglia following ocular infection. In order to identify the nucleotide sequences responsible for the differences in the capacity of the two HSV strains to grow within the peripheral nervous system, an intertypic recombinant was generated by infecting neuroblastoma cells with HSV-2(186) and a HSV strain possessing nucleotide sequences from HSV-1(17). The genome of the intertypic recombinant was composed entirely of HSV-2(186) DNA except for 2.0 kb of HSV-1(17) DNA positioned between m.u. 0.413 and 0.426. Following corneal infection of mice, the intertypic recombinant grew to higher titers in both ocular tissues and trigeminal ganglia than did the HSV-2 parent. Most significantly, the intertypic recombinant could spread into the brain from the trigeminal ganglion and kill the host whereas mice inoculated with the HSV-2(186) parent survived infection. The 2.0 kb of HSV-1(17) DNA inserted into the genome of the intertypic recombinant encodes the 5' terminus of the HSV-1 gene for DNA polymerase. Thus, the results suggest that the difference in the capacity of two HSV strains to replicate within the trigeminal ganglion of its host and to spread into the brain is determined by nucleotide sequences within the gene for DNA polymerase.

Animals↗
PubMed1987

Nucleotide sequences important in DNA replication are responsible for differences in the capacity of two herpes simplex virus strains to spread from cornea to central nervous system.

Two herpes simplex virus (HSV) intertypic recombinants were isolated with genomes composed entirely of HSV-2(186) nucleotide sequences except for a 6.0 kb segment of HSV-1(17) DNA positioned between 0.40 and 0.44 map units. Following corneal infection of mice, HSV-1(17) and the two intertypic recombinants spread from infected eyes into the central nervous system and induced a fatal encephalitis. Ocular infection with the HSV-2(186) parent did not lead to detectable amounts of virus in the brain, and none of the mice developed encephalitis. The 6.0 kb HSV-1(17) DNA inserted within the genome of the two intertypic recombinants contained nucleotide sequences involved in DNA replication. These include the HSV-1(17) oriL, the HSV-1(17) gene for DNA polymerase and portions of the HSV-1(17) gene coding for DNA-binding protein ICP8. Thus, our results indicate that the difference in the capacity of HSV-1(17) and HSV-2(186) to spread from the cornea into the CNS is determined solely by nucleotide sequences associated with DNA replication.

Animals↗
PubMed1987

Failure of intertypic recombinant constructed from HSV-1 x HSV-2 virulent parents to induce ocular pathology.

An intertypic recombinant constructed from HSV-1 x HSV-2 parents was isolated which failed to induce any overt ocular pathology when inoculated onto the sacrificed cornea of four-week-old SJL/J mice. During the 24-48 hour post-infection period there was transient virus replication but by day 3 the infectious titer in the eye had dropped by greater than or equal to 10(4)-fold, and little or no virus could be recovered thereafter. When immunosuppressed (600 r) mice were infected corneally, virus clearance was delayed several days but again no obvious ocular pathology was seen, and no mice died. By contrast, infection of the cornea with either parent was followed by virus replication and development of clinically apparent pathology which could progress to blinding stromal keratitis. The genome of the intertypic recombinant was analyzed by agarose gel electrophoresis of restriction endonuclease digests and found to consist entirely of HSV-1 DNA except for HSV-2 DNA sequences located between map units 0.10-0.16, 0.41-0.43, and 0.77-1.0. Potential explanations for the loss of virulence are discussed.

Animals↗
PubMed1987

Superiority of antibody versus delayed hypersensitivity in clearance of HSV-1 from eye.

The contribution that antibody and delayed type hypersensitivity (DTH) make in promoting HSV-1 clearance from the infected cornea was investigated. Balb/c mice were immunized intravenously or subcutaneously with an attenuated strain of HSV-1 to generate hosts which were antibody-producing DTH-tolerant or antibody-producing DTH-responsive. Anti-mu serum treated mice were likewise sensitized intravenously or subcutaneously to obtain hosts which were antibody depressed-DTH tolerant or antibody-depressed DTH-responsive. Eight days after sensitization, these four sensitized groups and unsensitized controls were infected on scarified corneas with a stromal keratitis inducing strain of HSV-1, and the extent of virus replication was determined 1, 3, and 7 days later. Very different results were obtained depending upon the host's immune status. Virus proliferated extensively (greater than 3-4 logs) in the eyes of nonimmune mice and antibody-depressed DTH-tolerant hosts during the first 3 days after infection. In striking contrast, HSV-1 could not be detected even 24 hr post challenge in antibody-producing DTH-tolerant mice. In fact, such mice cleared virus from the eye as efficiently as immunologically intact hosts. However, in mice with the reverse immune status, ie antibody-depressed DTH-responsive, virus growth was clearly evident (greater than 2-3 logs) during days 1-3, and only thereafter did complete clearance occur. These results indicate that in the sensitized host antibody is both independent of and significantly more effective than DTH in promoting HSV-1 eradication from the infected eye.

Animals↗
PubMed1986

Herpes simplex virus type 1 DNA sequences which direct spread of virus from cornea to central nervous system.

The virulence of a herpes simplex virus (HSV) intertypic recombinant possessing HSV-1 DNA sequences from map units 0.31 to 0.44 and HSV-2 sequences from map units 0 to 0.30 and 0.45 to 1.0 were compared with the virulence of the two parental strains. Following ocular inoculation, both the intertypic recombinant and the HSV-1 parent replicated at the infection site and spread to the peripheral and central nervous system (CNS) to produce fatal encephalitis. The HSV-2 parent also replicated at the infection site but failed to progress to the CNS. However, when inoculated intracerebrally, the HSV-2 strain was as lethal as the HSV-1 parent. Furthermore, the HSV-2 strain could produce thymidine kinase at 37 and 39 degrees in levels comparable to the HSV-1 strain. The results indicate that transfer of the HSV-1 DNA sequences imparted to the recombinant virus the necessary genetic information to spread from the cornea into the central nervous system.

Animals↗
PubMed1985

Resolution of HSV corneal infection in the absence of delayed-type hypersensitivity.

The role of delayed-type hypersensitivity (DTH) in the resolution of herpes simplex virus type 1 (HSV-1) ocular infection was examined. Infection of Balb/c mice on the sacrificed cornea with HSV-1 resulted in sensitization for DTH. This response, demonstrable by swelling of the ear following inoculation with ultraviolet-irradiated virus, was optimal 7 days postinfection. The reaction was immunologically specific and characterized histologically by a predominately mononuclear cell infiltrate. DTH responsiveness could be completely abrogated if the mice were inoculated intravenously with an attenuated strain of HSV-17 days before corneal infection. DTH-unresponsive mice were, nevertheless, resistant to corneal challenge with sublethal or lethal doses of HSV-1. Resistance was accompanied by a greater than 30-fold reduction in infectious virus in the eye 24 hr post challenge. A cellular infiltrate characteristic of a DTH response was not observed within the cornea during virus clearance. Tolerance was restricted to DTH, as antibodies to HSV antigens could be readily demonstrated 6-7 days after intravenous virus immunization. These antibodies may have contributed to the resistance observed. The results establish that neither a systemic nor local DTH response is required by the host to resist HSV-1 ocular infection.

Animals↗
PubMed1979

Uninhibited growth and metastases of herpes simplex virus-transformed cells in virus-sensitized hosts.

Four established tumour lines of hamster cells transformed by herpes simplex virus (HSV) but not shedding the virus were examined for continued expression of virus-associated antigens. Hamster or rabbits, appropriately immunized to the tumour cells, produced virus-neutralizing antibody. The serum titres were invariably low, suggesting that only small quantities of virion antigen were present in the cells. Hamster sensitized to HSV and resistant to virulent virus challenge did not reject low numbers of tumour cells, nor was the incidence of lung metastases significantly reduced. Virus-sensitized lymph-node cells, readily cytotoxic for HSV-infected hamster embryo fibroblasts, did not lyse any of three transformed lines tested in an 18-h 51chromium release test. Animals that had their tumour excised demonstrated no or only modest resistance to tumour rechallenge. Thus, virus-specific transplantation rejection antigen could not be detected in the HSV tumour lines although low levels of virus structural antigens were present.

Animals↗
PubMed1979

Cellular immune response to human cytomegalovirus. I. Lymphocyte transformation studies in the rabbit.

The lymphocyte transformation assay was used to monitor the cellular immune response of rabbits sensitized to cytomegalovirus (CMV). Peripheral blood lymphocytes from animals inoculated with purified virus were specifically stimulated by crude or twice-banded CMV. Blood cells from rabbits immunized to herpes simplex virus type 1 (HSV-1) responded to that antigen but not to CMV. Viable or heated CMV preparations stimulated unwashed blood cells as efficiently as washed cells. Furthermore, preincubation of stimulating antigen with anti-CMV serum did not prevent lymphocyte activation. Lymphoid cells readily stimulated by virus antigen were not stimulated by cells transformed by CMV or HSV-1.

Animals↗
PubMed1979

Inability of antiserum active in antibody-dependent cellular cytotoxicity and arming tests to protect against simian virus 40 tumor cell challenge.

Previous studies have shown that simian virus 40 (SV40) hamster tumor cells that were pretreated with antiserum from syngeneic hosts sensitized to SV40 were killed following exposure to nonsensitized spleen cells. In this study preincubation of nonadherent spleen or lymph node cells with SV40 antiserum rendered the cells specifically cytotoxic for the SV40-transformed fibroblasts. The capacity of a particular antiserum to "arm" (i.e., to be made specifically cytotoxic) was comparable with its ability to mediate antibody-dependent cellular cytotoxicity (ADCC). Experiments were performed to determine if the SV40 antiserum had prophylactic activity. Two hours after passive transfer of serum, cytotoxic effector cells could be demonstrated in the blood, spleen, and mesenteric lymph node, and the recipients' sera were active in ADCC tests. Nevertheless, such hosts were not resistant to challenge with small numbers of SV40 tumor cells that were given intradermally or intracardiacly, nor was tumor growth suppressed by addition of normal lymph node cells to the antiserum-pretreated tumor cell inoculum. Thus SV40 antiserum, active at high titer in ADCC and arming assays, did not prevent or delay growth of SV40 tumor isografts.

Animals↗