HSV antigens and HSV DNA in avascular and vascularized lesions of human herpes simplex keratitis

Holbach, Leonard M.; Font, Ramon L.; Baehr, Wolfgang; Pittler, Steven J

doi:10.3109/02713689109020359

Cited by 27 publications

(16 citation statements)

References 15 publications

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“…We devised a method to obtain DNA from archival paraffin-embedded sections (1-3). Simply boiling tissue sections in the presence of detergent sufficiently released the DNA for PCR analysis (30). DNA released in this manner was then assessed by PCR amplification with a pnrmer pair (149/150) that amplifies a 298-bp segment of the rd (Pdeb) gene (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Excess water around the sections was carefully blotted with absorbent paper, and the sections were scraped while still moist into a Microfuge tube. DNA was liberated from the sections by a rapid boiling method (30).…”

mentioning

confidence: 99%

“…PCR amplification was performed as described (30) except that denaturation, annealing, and extension steps were 30 sec each. DNA sequencing of PCR products was carried out essentially as described (29).…”

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confidence: 99%

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PCR analysis of DNA from 70-year-old sections of rodless retina demonstrates identity with the mouse rd defect.

Pittler

Keeler

Sidman

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

133

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Rodless retina (gene symbol, r) was discovered in mice by Keeler 70 years ago and was first described in this journal as an autosomal recessive mutation leading to "the absence of the visual cells (rods), the external nuclear layer, and the external molecular layer" [Keeler, C. E. (1924) Proc. Natl. Acad. Sci. USA 10, 329-333]. The mutation was studied by Keeler and others in the United States and Europe over the next decade, but Keeler's stock was destroyed in 1939, and mice dermitively related to his by pedigree and progeny tests also appeared to have been lost by the end of World War II. In the early 1950s Bruckner in Basel recognized mice with a similar retinal phenotype. Investigators in London and Strasbourg analyzed descendants of Bruckner's mice and concluded, on the basis of different pathogenesis from r, that they carried a new mutation, which came later to be called retinal degeneration, rd. The relationship of r and rd has been unsettled ever since. Now that the rd phenotype is known to be due to a nonsense mutation in the rod photoreceptor cGMP phosphodiesterase 3-subunit gene, we hoped to settle the question by direct analysis of r DNA. DNA was liberated from 70-yearold histological sections of +/r and rlr eyes, the only extant r DNA, and the regions encompassing the nonsense mutation amplified by the polymerase chain reaction (PCR). Sequence analysis of the PCR products revealed the presence of the same nonsense mutation and two intron polymorphisms in r DNA. PCR and direct sequence analysis of 11 strains of mice known to carry rd (or a similar allele) also revealed the presence of the nonsense mutation and the same intron polymorphisms. The fact that all r and rd mice contain an identical defect and intron polymorphisms in the phosphodiesterase (3subunit gene settles beyond reasonable doubt that a single mutation arising >70 years ago is now widely distributed through inbred mouse strains. Because of the extensive use of the name in publications of the past 40 years, we propose that the gene continue to be designated retinal degeneration, rd.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

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PCR analysis of DNA from 70-year-old sections of rodless retina demonstrates identity with the mouse rd defect.

Pittler

Keeler

Sidman

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

133

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“…Vários métodos incluindo a cultura viral (14)(15)(16)(17)(18)(19) , reação da cadeia da polimerase (19)(20)(21)(22)(23)(24)(25) , imuno-histoquímica (25)(26) , e hibridização in situ (23) têm sido usados para o diagnóstico do HSV tipo 1 em córneas humanas. O trabalho de El-Aal AM (3) mostrou que a cultura do vírus herpes simples foi positiva em 10 (20,8%) de 48 casos suspeitos de ceratite herpética epitelial dendrítica com infiltrado subendotelial.…”

Section: Discussionunclassified

Detecção do vírus herpes simples por reação em cadeia da polimerase em pacientes com ceratite herpética típica ou atípica

Kamimura

Takata

Fernandes

et al. 2008

Arq. Bras. Oftalmol.

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“…The mechanisms underlying the latency and reactivation of HSV-1 infection in humans are but poorly understood. The existence of herpetic DNA in the absence of manifested symptoms of its activity has been demonstrated in both normal and diseased corneal tissue in human [Rong et al, 1988;Crouse et al, 1990;Cantin et al, 1991;Holbach et al, 1991;Kaye et al, 1991;Ohashi et al, 1991;Rong et al, 1991] and animal studies [Openshaw, 1983;Cantin et al, 1992]. Viral antigen, presented to the immune system by corneal epithelial cells [Fahy et al, 1993], triggers a cytokineinduced overexpression of class I and II human leukocyte antigens in the central corneal epithelium and stroma [Pepose et al, 1985], where it attracts CD4-positive T cells and Langerhans cells [Hendricks et al, 1992].…”

Section: Discussionmentioning

confidence: 99%

Low rate shedding of HSV-1 DNA, but not of infectious virus from human donor corneae into culture media

Garweg

Boehnke

1997

J. Med. Virol.

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Fluid samples derived from 451 organ cultured corneae were tested for the presence of HSV-1 DNA after electroseparation and amplification for fragments of the glycoprotein D- and thymidine kinase-encoding genes. Of the culture media, 134 were processed immediately after withdrawal (Group 1); 100 were stored at ambient temperature for 6 to 60 weeks (Group 2); 90 were stored at -8 degrees C for 4 to 9 weeks (Group 3); and 127 were stored at -20 degrees C for 2 to 30 weeks (Group 4). The degradation of human DNA (marker gene, betaglobin) under these different storage conditions and of human and HSV-1 DNA as a sequential function of time at ambient temperature was gauged by the loss of a detectable signal for the respective component. Endothelial cell density within each of the corneal discs was determined before and after organ culture. In 7/451 culture fluid samples, HSV-1 DNA corresponding to either the glycoprotein D- or thymidine kinase-encoding genes was detected. In culture fluid samples derived from Group 2 at ambient temperature, for 6 to 60 weeks) and 3 (at -8 degrees C, for 4 to 9 weeks), complete degradation precluded the detection of human DNA, and hence probably also of HSV-1 DNA; only at -20 degrees C did DNA remain stable for protracted periods of time. Even so, HSV-1 DNA was detected in only 2% of those media in which no degradation was to be expected; additionally, there existed no correlation between its presence in culture fluid samples and the loss of endothelial cells or cytopathic changes. DNA can be extracted successfully and concentrated twenty-fold from high-volume samples by electroseparation. When shed into culture fluid, it is remarkably prone to a time and temperature dependent degradation, which may lead to false negative results. It is concluded that there is no infectious virus to be expected in the specimens; the occurrence of HSV-1 DNA in donor corneae would not appear to be an important factor influencing their biological quality during the period of organ culture.

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HSV antigens and HSV DNA in avascular and vascularized lesions of human herpes simplex keratitis

Cited by 27 publications

References 15 publications

PCR analysis of DNA from 70-year-old sections of rodless retina demonstrates identity with the mouse rd defect.

PCR analysis of DNA from 70-year-old sections of rodless retina demonstrates identity with the mouse rd defect.

Detecção do vírus herpes simples por reação em cadeia da polimerase em pacientes com ceratite herpética típica ou atípica

Low rate shedding of HSV-1 DNA, but not of infectious virus from human donor corneae into culture media

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