Utility of HSV Serology for Chronic Corneal Pathology.

OBJECTIVES: The purpose of this study is to determine the utility of obtaining herpes simplex virus (HSV) serology for patients presenting with chronic corneal pathology of unknown etiology for which HSV is a diagnostic consideration. METHODS: A retrospective analysis was performed of all patients presenting to one cornea specialist (J.M.G.) between August 2011 and April 2018 with a chronic (>6 weeks) corneal condition for which HSV was suspected and serology was performed. Patient demographics, clinical presentation, treatment, final diagnosis, and follow-up duration were recorded. RESULTS: Fifty-four patients with a median age of 52 (range: 5-85) years were included in the study. Patients were classified by presenting clinical features as corneal ulcer (46.2%), stromal keratitis (24.1%), superficial keratitis (18.5%), or keratouveitis (11.1%). The seroprevalence of HSV-1 and HSV-2 antibodies were 42.6% and 18.5%, including 5 patients (9.3%) positive for both HSV-1 and HSV-2. Serology impacted management for all patients with negative titers (48.1%), defined as discontinuing antiviral medication, electing not to start antiviral medication, or continuing antiviral medication for a non-HSV etiology (e.g., varicella zoster keratitis). No patients with HSV serology were ultimately diagnosed with HSV keratitis. Median follow-up duration was 1.5 years (range 0.8-6.6 years). CONCLUSION: Although only useful when negative, our study confirms that serology may be useful for excluding HSV as a diagnostic consideration for patients presenting with chronic corneal pathology. The seroprevalence of HSV antibodies for our patient cohort was comparable with previous population-based studies.

A crucial role for B and T lymphocyte attenuator in preventing the development of CD4+ T cell-mediated herpetic stromal keratitis.

PURPOSE: To investigate the effect of the B and T lymphocyte attenuator (BTLA; CD272) on cluster of differentiation (CD)4(+) T cell-mediated corneal immunopathology during murine herpetic stromal keratitis (HSK). METHODS: BALB/c mice were infected with the herpes simplex virus type 1 (HSV-1) KOS strain by corneal scarification. The levels of BTLA expression in CD4(+) and CD8(+) T cells in murine peripheral blood were determined by flow cytometry on days 0, 3, 7, 10, 14, and 21 after HSV-1 infection. BTLA expression in the infected cornea was detected by immunohistochemistry. BALB/c mice were injected intraperitoneally with recombinant plasmid DNA encoding BTLA (pBTLA), pcDNA3.1, or PBS on 0 and 7 days before infection and 7 days postinfection. The incidence and severity of stromal disease, tear film virus titers, and the delayed-type hypersensitivity (DTH) reaction were then compared among treated and control groups. The effects of pBTLA on CD4(+) T cells that infiltrated into infected corneas and on type 1 helper T-cell (Th1) cytokines (interferon-gamma [IFN-γ]) were evaluated. The levels of glycoprotein D (gD) mRNA in corneas were tested by real-time PCR. The eyes were examined histologically. RESULTS: BTLA expression increased both in the corneas of HSV-1 infected mice and in CD4(+) T cells in the murine peripheral blood. Systemic administration of pBTLA resulted in a diminished incidence and severity of corneal lesions compared to controls. Treatment with pBTLA led to a decreased infiltration of CD4(+) T cells into infected corneas, and diminished Th1 responses in murine corneas, draining lymph nodes, and splenocytes. The pBTLA treated mice showed an impaired DTH response two weeks after HSV-1 infection compared to control mice. No differences were noted in tear film virus titers or gD mRNA levels in corneas among the experimental groups. CONCLUSIONS: The results suggest that recombinant pBTLA plays a crucial role in preventing HSV-1 specific responses in CD4(+) Th1 cells in the infected corneas. Thus, BTLA, with immunosuppressive effects, may be a good candidate for treatment of HSK.

Involvement of STAT4 in IgG subtype switching and ocular HSV-1 replication in mice.

PURPOSE: To assess the relative impact of elevated T-helper 2 (T(H)2)- and reduced T-Helper 1 (T(H)1)-dependent immune responses on ocular herpes simplex virus type 1 (HSV-1) infection. METHODS: Signal transducer and activator of transcription protein 4 knockout mice (BALB/c-STAT4(-/-)) and wild-type BALB/c control mice were immunized with avirulent HSV-1 strain KOS or were mock-immunized. Three weeks after the third immunization, neutralizing antibody titers were determined by plaque reduction assays. Following ocular infection with virulent HSV-1 strain McKrae, viral replication in the eye, blepharitis, corneal scarring (CS), survival, and immunoglobulin (Ig) isotypes in sera were determined. RESULTS: Vaccinated STAT4(-/-) and BALB/c mice contained significant and similar neutralizing antibody titers and were completely protected against HSV-1-induced death and CS. In contrast to vaccinated STAT4(-/-) mice, mock-vaccinated STAT4(-/-) mice had higher ocular HSV-1 titers than mock-vaccinated BALB/c mice on days 2-3 post-ocular infection. There were also significant differences in the levels of IgG2a, IgG2b, and IgG3 in the sera of STAT4(-/-) mice when compared to the control BALB/c mice. CONCLUSIONS: These results suggest that the absence of T(H)1 cytokine responses did alter protection against viral replication and IgG isotypes but not eye disease or survival.

[Imbalance of alpha- and gamma-interferon levels in patients with herpetic keratitis].

The paper presents data on elevated serum and lacrimal fluid alpha- and gamma-interferon (INF) levels in patients with herpetic keratitis (HK) in the course of the disease depending on its clinical form and stage. A more significant increase in the levels of alpha- and gamma-INF was noted in the lacrimal fluid, which was indicative of the important role of local antiherpetic corneal and conjunctival protection. The determination of serum and lacrimal fluid IFN levels in patients with HK may be used to predict the clinical course of the disease.

[Use of testosterone preparations in complex therapy of severe herpetic keratitis in male patients]. / Primenenie preparatov testosterona v kompleksnoi terapii tiazhelykh form gerpeticheskogo keratita u lits muzhskogo pola.

Data are reported on the efficiency of using the testosterone preparations in male patients with severe herpetic keratitis concomitant with a reduced concentration of the hormone in blood serum; simultaneously, zovirax (an antiviral drug) and immunocorrector tymalin were administered. The above drugs normalized the testosterone content in blood serum in 46.9% of patients; they stabilize the count of CD95+ lymphocytes in peripheral blood, which is manifested clinically by an improved visual acuity and a shorter treatment of the above patients in hospital.

[Interaction of the hormonal and immunologic impairments in men with herpetic keratitis]. / Vzaimosviaz' gormonal'nykh i immunologicheskikh narushenii pri gerpeticheskom keratite u muzhchin.

Data are described in the paper on the significance of hormonal and immunological impairments and on their interrelation within the pathogenesis of herpetic keratitis in males. An evaluation of contents of testosterone, cortisol, CD8+, CD95+ lymphocytes, interleukin-6 and tumor necrosis factor-alpha (TNF-alpha) as well as definition of interrelations between them has a prognostic value for males with herpetic keratitis. A dropping concentration of testosterone as observed in males with severe herpetic keratitis is indicative of the feasibility to add some hormone-correcting drugs to the combined therapy.

[Cytokine status of patients with ophthalmic herpes]. / Tsitokinovyi status bol'nykh oftal'mogerpesom.

Data are reported on the content of pro- and antiinflammatory cytokines in blood serum and lachrymal fluid of 62 patients with ophthalmoherpes. It was concluded that the cytokine profiles are subject to changes in the eye herpes: the IL-6 and IL-4 contents were increasing in blood serum, while IL-1 beta and TNF-alpha were increasing in lachrymal fluid. The content of cytokine in blood serum is different from such content in lachrymal fluid in surface-type and profound variations of ophthalmoherpes. Disbalance of inflammatory IL-1 beta, IL-6, TNF-alpha and IL-4 are typical of the latter case, which can be used clinically to evaluate a disease severity degree.

[Activity of the antioxidant enzymes of erythrocytes in herpetic keratitis]. / Aktivnost' antioksidantnykh fermentov éritrotsitov pri gerpeticheskom keratite.

Free-radical oxidations get activated, the activity of catalase slows down and the activity of superoxiddismutase increases in peripheral blood erythrocytes of patients with herpetic keratitis, which is indicative of a changing antioxidant protection in late stages of herpetic infection. The results reveal new chains in the evolution of ophthalmic herpes and confirm the feasibility of adding antioxidants to the complex therapy of patients with herpetic keratitis.

Mapping of genes involved in murine herpes simplex virus keratitis: identification of genes and their modifiers.

Herpes simplex keratitis (HSK) is an inflammatory response to viral infection and self antigens in the cornea and is a major cause of blindness. Using two strains of mice which are susceptible (129/SVEV) and resistant (C57BL/6) to herpes simplex virus (HSV) strain KOS, (129/SVEV x C57BL/6)F(2) mice were generated and examined for their disease susceptibility in terms of clinical symptoms, ocular disease, and antibody production following corneal scarification with HSV (KOS). A genome-wide screen was carried out using microsatellite markers to determine the genetic loci involved in this response. Loci on chromosomes 4, 5, 12, 13, and 14 were shown to be involved in general susceptibility to clinical disease, whereas loci on chromosomes 10 and 17 were shown to be unique to ocular disease.

[Time course of interferon status of patients with herpetic keratitis during treatment with actipol, a new inteferon inducer]. / Dinamika interferonogo statusa u bol'nykh s gerpeticheskimi keratitami pri lechenii novym induktorom interferona aktipolom.

Interferon (IFN) status was evaluated in 20 patients with various forms of herpetic keratitis over the course of treatment with actipol (0.07% para-aminobenzoic acid). Actipol was injected subconjunctivally parabulbarly in combination with instillations into the conjunctival sac or only instilled, depending on the disease severity. The following parameters were evaluated: 1) total content of various IFN types in circulating blood and 2) leukocyte capacity to produce IFN in vitro in patient's whole blood cells. IFN-alpha was induced with Newcastle disease virus and IFN-beta with staphylococcal enterotoxin. Before treatment IFN-producing capacity of blood cells in vitro was decreased in 60% patients and was normal or increased in 40%. Plasma concentrations of IFN were moderately increased in 45% patients. All IFN parameters in patients with initially disordered IFN status normalized during actipol therapy and after clinical cure and did not change in the patients with initially normal IFN values. Hence, local actipol therapy in patients with various forms of ophthalmic herpes modifies IFN status.

[Analysis of corneal pathologic changes and laboratory parameters in herpes simplex karatitis patients with ganhuo-shangyanzheng or ganshengyinxuzheng].

In order to investigate the relationship between Ganhuo shangyanzheng and Can-shengyin xuzheng in herpes simplex karatitis patients, we observed corneal pathologic changes and examined blood levels of prostaglandin F2 (PGF2), prostaglandin E2 (PGF2), tumor necrosis factor (TNF), arginine vasopressin (AVP), norepinephrine (NR), epinephrine (E) in sixty herpes simplex karatitis patients with Ganhou shangyanzheng or Gan-shengyinxuzheng. The results showed that the corneal pathologic changes were corneal ulcer infiltrating to stroma of cornea in Ganhuo shangyanzheng patients, and refractary corneal ulcer with large amount of corneal neovascularizaton and infiltration of corneal stroma in Gan-shengyinxuzheng patients, the blood levels of PGF2, PGE2, TNF, AVP, NE, E in Ganhuo shangyanzheng patients were higher than those in Gan-shengyinxuzheng patients or healthy persons. The results suggest that these parameters may be objective parameters for differential diagnosis of traditional Chinese medicine Zheng types in patients with herpes simplex karatitis.

[Mechanisms for the therapeutic effect of herpes polyvaccines in chronic ophthalmic herpes and genital herpes]. / Mekhanizm lechebnogo éffekta gerpeticheskoi polivaktsiny pri khronicheskom oftal'mogerpese i gerpes genitalii.

Commercial inactivated culture polyvaccine against herpes simplex viruses (types 1 and 2) developed at D. I. Ivanovsky Institute of Virology promoted cessation of viremia. During the first vaccination viremia coincided with appearance of a focal allergic test on the retina, which is proposed for the diagnosis of herpetic involvement of the posterior compartment of the eye. T-cellular immunity normalized after a course of vaccination. Experimental immunization of rats and vaccination of patients with chronic ophthalmic and genital herpes demonstrated the therapeutic activity of inactivated herpetic polyvaccine in suppositoria.

[The expression of human leukocyte antigen from peripheral blood of patients with herpetic keratitis].

PURPOSE: To understand the correlation between the expression of human leukocyte antigen from peripheral blood and herpetic keratitis so as to find the pathogenesis of HSK. METHODS: The expression of HLA-ABC, HLA-DR, HLA-DP, HLA-DQ from peripheral blood of 32 cases with herpes simplex keratitis (the stromatic and ulcer herpetic keratitis are 16 cases respectively), comparing with the normal people, were investigated by flow cytometry. RESULTS: The expression rate of HLA-ABC from peripheral blood of patients with herpetic keratitis (both the stromatic and ulcer groups) was lower, and the expression of HLA-DR and HLA-DP was higher than normal people. No difference was found between the stromatic and the ulcer herpetic keratitis in the expression of HLA-ABC, HLA-DR, HLA-DP, HLA-DQ. CONCLUSION: The immune model in patients with herpetic keratitis is different from normal people. The individual with this kind of immune model can not destroy the infected virus effectively, also show autoimmunization to cornea.

Dosing considerations for oral acyclovir following neonatal herpes disease.

Herpes simplex virus lesions recur in 8-30% of infants who receive a course of parenteral antiviral therapy for an initial infection. Long-term acyclovir is used by some clinicians to prevent recurrent Herpes simplex disease. We describe nine infants who were treated with doses of oral acyclovir which were chosen to achieve 2-h post-plasma concentrations of > or = 2 micrograms/ml. Eight infants had Herpes simplex encephalitis and one had multiple recurrences of dermal and ocular disease. The target plasma concentration was chosen in order to attain acyclovir cerebrospinal fluid distribution (< or = 50% plasma) for an estimated ID30 of Herpes simplex II strains of 0.1-0.5 microgram/ml. One of nine patients failed to achieve the target plasma acyclovir concentration. One of nine patients developed symptomatic recurrence of the central nervous system disease and none of the remaining eight patients experienced recognized dermal or neurologic recurrence of Herpes simplex disease. Renal and neurologic status were routinely monitored and no signs of acyclovir toxicity were observed. Plasma concentration of acyclovir > or = 2 micrograms/ml may be achieved with average oral doses of 1340 mg/m2/dose (1000-1740 mg/m2/dose) given at 12-h intervals.

[Dyslipidemia in herpetic infection]. / Dislipidemiia pri gerpeticheskoi infektsii.

Herpes ophthalmicus (HO) patients were examined for lipid spectrum of the serum. The tests revealed dyslipidemia (DLE) with a distinctive rise in the levels of total cholesterol (CS), alpha-CS and beta-CS. The severity of the lipidemic shifts correlated with that of the infection. DLE was more marked in recurrent HO. Clinical evidence was consistent with experimental findings. In rabbit models, herpetic keratoconjunctivitis was characterized by pronounced lipidemic alterations correctable with antiherpetic drug furavir. The results are discussed in terms of herpetic infection pathogenesis and its role in the development of atherosclerosis.

Prophylactic acyclovir effectively reduces herpes simplex virus type 1 reactivation after exposure of latently infected mice to ultraviolet B.

PURPOSE: To determine the potential efficacy and anatomic sites of action of prophylactic oral acyclovir using a murine model of ultraviolet-B-induced reactivation of herpes simplex 1 keratitis. METHODS: Latent infection with herpes simplex 1 (McKrae) was established in 80 National Institutes of Health inbred strain of mice. Forty of the mice were given acyclovir orally and the other 40 latently infected mice served as controls. Mice were exposed to 250 mJ/cm2 of ultraviolet-B radiation and killed on days 1, 2, 3, and 4 after ultraviolet-B radiation. Trigeminal ganglia and eyes from these mice were homogenized and incubated on Vero cell monolayers for recovery of reactivated virus. RESULTS: Based on the recovery of infectious virus after ultraviolet-B in treated versus control groups, acyclovir effectively reduced detectable viral reactivation at both the ocular level (P = 0.003) and the ganglionic level (P = 0.025). The numbers of viral culture-positive eye and trigeminal ganglia homogenates in the control group were 11 and 6 out of 40, respectively, compared to 1 and 0 out of 40 culture-positive eye and trigeminal ganglia homogenates in the acyclovir treated mice. Therapeutic serum levels of acyclovir were confirmed by high performance liquid chromatography. In the acyclovir-tested group, the single case of viral break-through at the ocular surface was not an acyclovir-resistant mutant. CONCLUSION: Prophylactic acyclovir effectively reduces the incidence of herpes simplex virus-1 reactivation after ultraviolet-B-induced reactivation in National Institutes of Health inbred strain of mice.

MHC II-restricted, CD4+ cytotoxic T lymphocytes specific for herpes simplex virus-1: implications for the development of herpetic stromal keratitis in mice.

Herpetic stromal keratitis (HSK) appears to represent an immunopathological reaction in which CD4+ T cells play a prominent role. However, the exact immunopathological mechanism(s) utilized by CD4+ T cells during HSK remains to be elucidated. In this study, the presence of cytotoxic CD4+ T lymphocytes in the cervical and retropharyngeal lymph nodes of Balb/c mice experiencing HSK was investigated. After in vitro depletion of CD4+ or CD8+ T cells with specific monoclonal antibodies and complement treatment, the cytotoxic functions of the remaining T cell populations were assayed by using target cells expressing either MHC Class I or both Class I and Class II. Our results showed the presence of a distinct cytotoxic T lymphocyte (CTL) population which was CD4+ and demonstrated lytic activity in a Class II-restricted fashion. Furthermore, these cells were able to develop into efficient effector CTL in the absence of CD8+ T lymphocytes as assessed by in vivo depletion experiments. Immunohistochemical methods were also utilized to show the presence of both CD4+ lymphocytes and I-A+ cells in the corneal tissues during HSK. These findings support the notion that direct lysis of infected Class II-bearing corneal cells by CD4+ CTL might be one of the mechanisms leading to stromal immunopathology in herpetic infections.