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Biomedical subjects

B T Rouse

Publications and source records attributed to B T Rouse.

At least 37 records · Page 2Linked to original sources
PubMed2000

Pathogenesis of herpes simplex virus-induced ocular immunoinflammatory lesions in B-cell-deficient mice.

The role of B cells and humoral immunity in herpes simplex virus (HSV) ocular infections was studied in immunoglobulin mu chain gene-targeted B-cell-deficient mice (muK/O). At doses of virus well tolerated by immunocompetent mice, heightened susceptibility of muK/O mice to herpetic encephalitis as well as to herpetic stromal keratitis (HSK) was observed. An explanation was sought for the increased severity of HSK in the muK/O mice. First, the lack of antibody responses in muK/O mice resulted in longer viral persistence and dissemination to the corneal stroma, the site of inflammation. Prolonged virus expression in the corneal stroma was suggested to cause bystander activation of Th1-type CD4(+) T cells, further contributing to the severity of HSK lesion expression in muK/O mice. Second, muK/O mice generated minimal Th2 cytokine responses compared to wild-type mice. Such responses might serve to downregulate the severity of Th1-mediated HSK lesions.

Animals↗
PubMed2000

Identification of select lymphocyte homing molecules and vascular addressins in lymphotoxin-alpha deficient mice.

The transmigration of lymphocytes across vascular endothelium is a critical step for the localization of lymphocytes to lymph nodes in both naive and immune reactive states. Mice deficient in lymphotoxin-alpha (LT-alpha) lack peripheral and gut associated lymph nodes. Lymphocyte function and homing ability are reported to be normal in these mice yet information regarding cell adhesion molecules and counterpart vascular addressins is lacking. The phenotype of peripheral lymphocytes from LT-alpha deficient mice was investigated by the use of fluorescent activated cell sorting and immunohistochemistry. No difference was detected in the splenocyte and tissue expression of L-selectin, alpha4beta7 or its individual integrin components, mucosal addressin cell adhesion molecule (MAdCAM-1), intracellular adhesion molecule (ICAM-1), peripheral node addressin (PNAd), or platelet/endothelial cell adhesion molecule (PECAM-1) between wild-type and LT-alpha deficient mice. Therefore, impaired expression of these lymphocyte homing and vascular addressin molecules is apparently not included in the phenotype of the LT-alpha deficient mouse.

Animals↗
PubMed2000

Bystander activation of CD4+ T cells accounts for herpetic ocular lesions.

PURPOSE: Stromal keratitis is an immunopathologic consequence of herpes simplex virus (HSV) infection of the cornea. The lesion is immunopathologic, but the identities of molecules that drive the reaction remain unresolved. To exclude viral antigen recognition as a necessary step in the disease process, ocular HSV infection was followed in Tg-RAG mice (OVA-TCR transgenic mice crossed to RAG2-deficient mice) whose limited T-cell repertoire did not include immune responsiveness to HSV. METHODS: Mice with T-cell specificity to OVA peptide (Tg-RAG mice) as well as control DO11.10 and BALB/c mice were infected with HSV on the scarified cornea and subjected to clinical, histologic, and immunologic analysis. To evaluate involvement of OVA-specific CD4+ T cells in lesion development in Tg-RAG mice, monoclonal antibody to CD4+ T cells was used for in vivo CD4+ T-cell depletion. RESULTS: Tg-RAG mice were capable of eliciting ocular lesions in the absence of detectable reactivity to viral antigens. Lesion manifestation in Tg-RAG mice was CD4+ T-cell dependent and the cellular infiltrates and their inflammatory products in the HSV-infected cornea were comparable to similarly infected BALB/c and DO11.10 mice. CONCLUSIONS: The authors conclude that mechanisms other than viral antigen recognition, and hence molecular mimicry, are at play and are sufficient to cause HSV-induced stromal keratitis. The data imply a significant role for non-virus-specific CD4+ T cells that could become activated by an inflammatory milieu consisting of enhanced accessory molecules and proinflammatory cytokines in the cornea.

Animals↗
PubMed1999

Distribution fate and mechanism of immune modulation following mucosal delivery of plasmid DNA encoding IL-10.

DNA vaccination has been widely studied in several models of vaccination and in the treatment of inflammatory diseases, even though the mechanism involved is still unclear. This report demonstrates that mucosal administration of plasmid DNA leads to rapid and widespread distribution around the body. Dissemination likely occurred via the bloodstream because plasmid DNA was present in blood plasma. The plasmid DNA was also detectable in several tissues including draining lymph nodes, spleen, liver, bone marrow, and even the dermis of ear pinnae. Except for the site of administration, plasmid DNA was no longer detectable in tissues after 3 wk postadministration. RNA and protein expression was also found in the tissues and bloodstream. Animals previously primed by HSV infection and subsequently given IL-10 DNA via the nasal mucosa, showed diminished Ag-induced delayed type hypersensitivity reactions for up to 5 wk posttreatment. The mechanism of modulation involved diminished the Ag-specific proliferation and production of Th1 cytokines. The Ag-specific silencing effects persisted beyond the duration of detectable plasmid encoded protein and was maintained upon adoptive transfer of T cells into a plasmid-free environment. The silenced T cells were not a source of IL-10, and their anergic state was reversible by exposure to Ag in the presence of exogenous IL-2.

Adjuvants, Immunologic↗
PubMed1999

Immune modulation by IL-10 gene transfer via viral vector and plasmid DNA: implication for gene therapy.

In the present report, we have evaluated and compared the modulatory effect of the cytokine interleukin (IL)-10 expressed via viral vector or plasmid DNA on viral antigen-induced cutaneous inflammatory lesions. Our data demonstrate the superior potency of both recombinant vaccinia virus and herpes simplex virus IL-10 expression vectors after single intramuscular administration, but the effects were only short term and only functioned in animals lacking immunity to the viral vectors used for modulation. In contrast, modulatory effects achieved by plasmid DNA expressing IL-10 were delayed in onset and milder in effect but were far more persistent than those achieved by viral vectors. Moreover, plasmid DNA expressing IL-10 provided effective modulation when given repeatedly to animals. Our data also showed that IL-10 gene delivery resulted in a systemic and durable modulatory effect while the effect caused by a single IL-10 protein treatment was transient and confined to the injected site. Our results imply that the viral vector system is superior for obtaining short-term effects, whereas the plasmid DNA approach represents a better strategy to achieve gene therapy to modulate chronic inflammatory lesions.

Animals↗
PubMed1999

Resistance to herpetic stromal keratitis in immunized B-cell-deficient mice.

This study evaluates the role of antibody as an indicator of immunity to ocular challenge with herpes simplex virus (HSV). Two genotypes of mice, BALB/c or BALB/c with mu-chain knockout (muK/O; which lack functional B cells), were immunized systemically either with nonvirulent infectious virus or with a eukaryotic expression plasmid encoding glycoprotein B (gB). Whereas naive muK/O mice were 10- to 100-fold more susceptible to HSV infection than BALB/c mice, following immunization both groups showed similar levels of resistance to ocular challenge. Thus both HSV-immunized groups cleared virus within 3 days and showed no signs of ocular lesions. gB DNA-immunized mice cleared virus less rapidly (5 days), and the incidence of lesions was 10 and 25% in BALB/c and muK/O mice, respectively. Since muK/O mice failed to produce detectable anti-HSV antibody, the mechanism of rapid viral removal was assumed to have a T cell basis. However, T cells would likely not mediate any protection directly since such cells were absent in infected corneas during clearance. A likely mechanism of immunity could involve innate defenses, perhaps enhanced by the action of cytokines released from antigen-reactive CD4+ cells in vascularized tissue adjacent to the cornea. Thus an abrupt inflammatory response consisting principally of neutrophils occurred in the corneal stroma in immune mice, and this subsided when virus disappeared. These data reveal that even though the deficiency in generating antibody renders mice more susceptible to HSV infection, once primed, resistance to disease expression is mediated solely by the cellular components and their products.

Animals↗
PubMed1999

Bystander activation of CD4(+) T cells can represent an exclusive means of immunopathology in a virus infection.

Herpetic stromal keratitis (HSK) is an immunopathological lesion involving herpes simplex virus (HSV) infection and CD4(+) T cells of the Th1 phenotype, but the nature of the target antigens which drive HSK remains uncertain. In the present report we show that ovalbumin TCR-transgenic mice backcrossed to SCID mice unable to recognize HSV show clinical signs of HSK but die of viral encephalitis before the lesions become severe. However, passive transfer of anti-HSV serum at 24 h clears virus and affords protection from both HSK lesions and death. Adoptive transfer of CD8(+) T cells at 72 h usually conferred protection but animals developed severe corneal pathology by 3 weeks post infection. At this time viral antigens were not demonstrable in the cornea and the T cells in the inflammatory lesions were CD4(+)KJ1-26.1 idiotype positive, i. e. OVA peptide specific. These results indicate bystander activation of CD4(+) T cells in a virus-induced inflammatory milieu. This mechanism of immunoinflammation may represent an important component of any lesion which involves CD4(+) T cells.

Adoptive Transfer↗
PubMed1999

Chemokines and ocular pathology caused by corneal infection with herpes simplex virus.

The role played by chemokines in disease process is an active area of research that continues to uncover new players. In this report we discuss the likely role of selected chemokines in the disease herpetic stromal keratitis (HSK). This lesion occurs as a sequel to herpes simplex virus infection and is currently accepted as an immunopathological process which primarily involves CD4+ T lymphocytes. In this review we discuss the events involved in HSK, the chemokine profile associated with this disease, and speculate on cellular activities and molecular events which characterize HSK as an immunopathological disease.

Animals↗
PubMed1998

Immune responsiveness of lymphotoxin-alpha-deficient mice: two reconstitution models.

The effects of lymphotoxin-alpha (LT-alpha) deficiency on mucosal immune status has not been defined. We utilized severe combined immunodeficiency (scid) mice as recipients of both mutant and wild-type whole splenocytes to determine whether lymphocytes from mutant mice had impaired homing ability. We also utilized irradiated mutant mice as recipients of wild-type whole splenocytes to determine whether lymphoid tissue anlages had, indeed, failed to develop as a consequence of LT-alpha deficiency. Subsequently, all mice were immunized orally with an attenuated strain of Salmonella typhimurium and mucosal IgA responses were monitored. The data presented here demonstrate that scid recipients generate mucosal responses equally well when reconstituted with mutant or wild-type lymphocytes. In contrast, reconstitution of mutant mice with wild-type cells failed to affect the efficiency of their mucosal immunity. The mutant phenotype, therefore, appears to involve neither impaired lymphocyte homing nor function in the generation of mucosal immunity. However, the mutant phenotype and immune responsiveness cannot be transformed merely by the provision of LT-alpha-expressing donor cell populations. The consequence of LT-alpha deficiency on mucosal immune responsiveness appears to be due to the lack of gut-associated lymphoid tissues, which may include the spleen, in mutant mice.

Animals↗
PubMed1998

Virus-induced immunoinflammatory lesions in the absence of viral antigen recognition.

Herpetic stromal keratitis (HSK) is a CD4+ T cell-controlled immunopathologic lesion in the eye that results from infection with herpes simplex virus (HSV). Target Ags involved in HSK remain undefined. In this study, we determined if HSK could be induced in animals genetically incapable of generating HSV Ag-specific CD4+ T cells. Mice bearing transgenic TCR specific to OVA peptide 323-339 (DO11.10) were crossed to SCID mice whose offspring (Tg-SCID) possessed CD4+ T cells, >98% of which expressed the OVA peptide-specific TCR. HSV infection of Tg-SCID mice was lethal, and mice failed to generate detectable T cell responses even after repeated immunization with a mutant avirulent virus (AN-1). Immunization with AN-1 virus followed by ocular challenge with HSV resulted in ocular inflammation before encephalitis, in contrast to the protection conferred in the control BALB/c and DO11.10 mice. These results indicate that clinical HSK may not require viral Ag recognition by CD4+ T cells and that T cells of irrelevant specificity can be recruited, activated, and driven into effector function in the HSV-infected cornea. This is suggested to represent a bystander activation effect resulting from the presence of proinflammatory mediators resulting from HSV replication.

Animals↗
PubMed1998

Immunomodulation by mucosal gene transfer using TGF-beta DNA.

This report evaluates the efficacy of DNA encoding TGF-beta administered mucosally to suppress immunity and modulate the immunoinflammatory response to herpes simplex virus (HSV) infection. A single intranasal administration of an eukaryotic expression vector encoding TGF-beta1 led to expression in the lung and lymphoid tissue. T cell-mediated immune responses to HSV infection were suppressed with this effect persisting as measured by the delayed-type hypersensitivity reaction for at least 7 wk. Treated animals were more susceptible to systemic infection with HSV. Multiple prophylactic mucosal administrations of TGF-beta DNA also suppressed the severity of ocular lesions caused by HSV infection, although no effects on this immunoinflammatory response were evident after therapeutic treatment with TGF-beta DNA. Our results demonstrate that the direct mucosal gene transfer of immunomodulatory cytokines provides a convenient means of modulating immunity and influencing the expression of inflammatory disorders.

Administration, Intranasal↗
PubMed1998

Role of mucosal immunity in herpes simplex virus infection.

This study evaluates whether the vaginal mucosal surface of immunized mice can prevent invasion by herpes simplex virus (HSV) and aims to identify immune components that affect immunity after challenge at the vaginal mucosa. Despite the induction of both IgA and IgG vaginal Ab following immunization with recombinant vaccinia virus vectors expressing either glycoproteins B or D, viral infection occurred in most animals even after minimal viral dose challenge. Challenged immune animals, including those genetically unable to generate anti-HSV Ab, survived and showed few if any clinical signs of infection. Experiments with T cell subtype knockout animals and depletion with T cell subset-specific MAb indicated that immunity following vaginal challenge was principally dependent on the function of CD4+ T cells. Our results indicate that anti-HSV vaccines may not provide barrier immunity at the vaginal mucosal site but may be adequate to minimize clinical expression of disease.

Administration, Intranasal↗
PubMed1998

Immunopathology of herpetic stromal keratitis: discordance in CD4+ T cell function between euthymic host and reconstituted SCID recipients.

Infection of the mouse cornea with herpes simplex virus (HSV) results in an immunopathologic disease of the eye termed herpetic stromal keratitis (HSK), in which the principal orchestrator is the CD4+ T cell. The mouse genotype largely determines susceptibility or resistance to HSK. BALB/c mice (H2dIgh-1a) are susceptible, while its congenic C.B-17 strain (H2dIgh-1b), which differs only in the Ig heavy chain locus, is resistant to HSK. As the magnitude and duration of viral replication as well as anti-HSV immune responses were similar in both strains, it was determined whether resistance was due to failure of CD4+ T cells to organize the immunopathologic reaction. Adoptive transfer of HSV-primed or naive CD4+ T cells from resistant C.B-17 strain into HSV-infected SCID mice resulted in HSK lesions indistinguishable from those caused by similar transfers of BALB/c CD4+ T cells. Similar results were obtained with transfers of whole T cell populations as well as with unfractionated splenocytes from the resistant mice. These results show that while intact C.B-17 mice exhibit resistance to HSK, they possess potentially pathogenic CD4+ T cells in their repertoire. The data suggest that the HSV-infected SCID mouse provides a proinflammatory microenvironment that overrides regulatory controls and/or cause activation of quiescent cells into aggressive effector T cells that orchestrate HSK.

Adoptive Transfer↗
PubMed1998

Modulation of mucosal and systemic immunity by enteric administration of nonreplicating herpes simplex virus expressing cytokines.

In this report the ability of enteric immunization with recombinant replication deficient (ICP4-/-) HSV expressing IFN gamma to generate protection and modulate mucosal and systemic immunity was evaluated. ICP4-/-HSV, ICP4-/-HSV expressing IL4, live replicating, and uv HSV were used as controls. Following enteric administration of live HSV, a Th1 cytokine response was induced in the spleen, while both Th1 and notable Th2 cytokine production were detected at mucosal sites. Modulation of mucosal and systemic immune response was achieved when nonreplicating recombinant HSV viruses expressing cytokines were used. Compared to the control replication defective viruses, decreased frequency of Th2 cytokine producing cells in Peyer's patches was observed following enteric administration of nonreplicating HSV expressing IFN gamma. When IFN gamma expressing virus was given enterically, modulation was observed at the systemic level, measured by ELISPOT for cytokine producing cells, ELISA from the in vitro restimulated splenic cell cultures, and by the increase of the IgG2a/IgG1 ratio in the serum. This report provides evidence that replication defective viruses expressing cytokine genes in contrast to uv HSV, are immunogenic when administered enterically and can generate significant immunomodulatory effects at the mucosal and systemic levels.

Animals↗
PubMed1998

Control of herpetic stromal keratitis using CTLA4Ig fusion protein.

Herpetic stromal keratitis (HSK) is an immunoinflammatory lesion in the cornea of the eye set off by infection with herpes simplex virus (HSV). The disease appears to be orchestrated by CD4+ T cells of the Th1 phenotype but the identity of target antigens involved in HSK remains unknown. In this proposal, we investigated if the inhibition of T cell activation with the fusion protein CTLA4Ig would abrogate the disease process when administered systemically. BALB/c mice infected with HSV-1 (RE strain) by corneal scarification were injected intraperitoneally on a single occasion with CTLA4Ig or L6 control (IgG Fc) given on day 2, day 5, or day 8 postinfection. Lesions in CTLA4Ig-treated mice showed markedly reduced severity judged by both slit lamp biomicroscopy and histopathology if treated on day 2 or day 5. Treated animals also expressed minimal HSV-specific splenic T cell and humoral antibody responses. Judged by the profile of T cell and IgG subset responses, inhibition by CTLA4Ig appeared more directly on the HSV-specific Th1 response, correlating with the known role of such cells in HSK. Delay of treatment until the time of disease onset (day 8) had marginal or negligible effects. The results indicate that blockade of coreceptor interaction between T cells and antigen-presenting cells during the induction phase of immune response significantly impairs onset and severity of herpetic stromal keratitis.

Abatacept↗
PubMed1998

The spleen and organized lymph nodes are not essential for the development of gut-induced mucosal immune responses in lymphotoxin-alpha deficient mice.

Lymphotoxin knock-out (KO) mice generate specific immune responses to orally administered immunogens despite having neither gut-associated nor peripheral lymphoid tissues. The spleen, therefore, was expected to play a role in the generation of immune responses in these KO mice. KO and wild-type (wt) mice were splenectomized and orally immunized with Salmonella typhimurium. Splenectomy produced the most profound effects on serum and fecal IgA levels in KO mice. Total and antigen-specific serum and fecal IgA were increased in splenectomized wt mice but decreased in splenectomized KO mice. Antigen-specific serum IgG was decreased in both KO and wt splenectomized mice while total IgG increased in splenectomized wt mice. Both splenectomized wt and KO mice demonstrated a compensatory expansion of the lamina propria compartment characterized by a significant increase in the number of IgA spot-forming cells. KO mice demonstrated further compensation for the loss of the spleen in the accelerated development of ectopic lymphoid tissues. We conclude that the spleen plays a prominent role as a lymphoid organ in KO mice but its removal does not abolish immune responsiveness. Residual immune responsiveness in splenectomized KO mice following oral immunization appears to be due to expansion and/or development of alternate effector compartments.

Animals↗
PubMed1998

Production of key molecules by ocular neutrophils early after herpetic infection of the cornea.

Herpes simplex virus infection of the eye can result in a blinding inflammatory lesion that is a T cell mediated immunopathological reaction. A prominent early event following HSV infection is neutrophil invasion of the corneal stroma. These cells may be involved in viral clearance and may influence the nature of the anti HSV T cell response which subsequently occurs. This article measures the expression of some key molecules which could participate in viral clearance and immune modulation. Using RT-PCR and in-situ hybridization, both corneal and peritoneal neutrophils were shown to be sources of iNOS and TNF alpha molecules which likely contribute to antiviral activity. Neutrophils also produce the cytokine IL-12, a key molecule which modulates the CD4+ T cell response to a type which mediates immunopathology. The present results indicate that neutrophils play an important role in the pathogenesis of herpetic ocular lesions.

Animals↗