Secondary Heterotypic Versus Homotypic Infection by Coxsackie B Group Viruses: Impact on Early and Late Histopathological Lesions and Virus Genome Prominence

doi:10.1016/S1054-8807(98)00025-8

Cardiovascular Pathology

Volume 8, Issue 2, March–April 1999, Pages 93-102

https://doi.org/10.1016/S1054-8807(98)00025-8 Get rights and content

Abstract

The impact of prior exposure to a different or identical strain of Coxsackievirus B (CVB) on murine CVB myocarditis was studied using a susceptible murine host (A/J[H-2 ^a]) and myocarditic CVB3 or avirulent CVB2 as primary or secondary infectants. The effects of secondary heterotypic infection (CVB2 followed by CVB3) and homotypic infection (CVB3 followed by CVB3) 28 days after primary inoculation, versus CVB2 or CVB3 alone, on injury and viral genomic replication, both early (day 7) and late (days 28 and 56), were evaluated. After the primary infection by CVB2, trivial viral RNA was present in the heart and other organs, and a substantial positivity was observed with CVB3 infection. Seven days after secondary heterotypic (CVB2-CVB3) infection, the quantity of CVB genome in heart, pancreas, liver, and spleen was increased compared with the virus genome in the CVB3-CVB3 group and in the group with primary CVB3 infection alone. This phenomenon was seen in the heart and spleen up to day 28 postsecondary infection. Tissue inflammation and necrosis in heart and pancreas were prominent 7 days postsecondary infection with CVB2-CVB3 and correlated well with an increased quantity of CVB genome. Virus genome was present in heart and spleen 28 days after CVB3 infection alone. Serum CVB3 neutralization titer was increased to 1:128 in CVB2-CVB3 group at days 7 and 28 postsecondary infection, and serum completely neutralized cytopathological effects of CVB3 in the CVB3-CVB3 group at day 7 and 28 postsecondary infection. Our results indicate that secondary heterotypic infection by CVB causes increased injury, inflammation, and CVB replication in target organs such as the heart and pancreas, as well as in immune compartments like the spleen. Compared with CVB3 alone, the intense inflammatory infiltrate in the CVB2-CVB3 group is as not due solely to postviral sensitization of the immune system, but rather to the inability of the host to eradicate the virus.

Section snippets

Mice and Virus Inoculation

Four-week-old male A/J[H-2^a] mice (Jackson Laboratory, Bar Harbor, ME) were housed in the Animal Resources Facility of the Max Planck Institut für Biochemie at Martinsried, Germany, and given laboratory mouse chow and water ad libitum.

The coxsackievirus B3_m (CVB3) RK variant, used in these experiments has been full-length sequenced (5). The coxsackievirus B2 (CVB2), Ohio-1 strain was obtained from the American Type Culture Collection (ATCC, Rockville, MD). Viruses were propagated in HeLa cell

Virus Genome Quantitation

Seven days after CVB2 infection, the quantity of CVB virus genome was not significantly different from sham-infected groups (p < .05) Figure 1, Figure 2, Figure 3, Figure 4. Because of the high degree of genetic identity shared among nonstructural gene regions of the human enterovirus group (9), detection of CVB2 and B3 genome is possible in a single hybridization assay. Consistent with our previous data, the quantity of CVB virus genome was significantly increased in heart, pancreas, liver,

Discussion

In the present study, CVB3-infected A/J mice developed extensive myocarditis at day 7 postinfection, with high subsequent morbidity. Histopathological examination of the infected myocardium revealed multiple foci of cardiomyocyte necrosis, predominant over, and not necessarily accompanied by, an inflammatory infiltrate. The strong correlation between histopathological scores and quantity of CVB genome early postinfection supports the concept that the cytolytic and necrotizing consequences in

Acknowledgements

The authors thank Mr. Lobus Bohunek, Mrs. Agripina Suarez, and Mrs. Helga Rieseman for their technical support. This work was supported by a Research Fellowship from the Heart and Stroke Foundation of Canada (JZY) and, in part, by an operating grant from the Heart and Stroke Foundation of British Columbia and Yukon (BMM).

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And the myocardia inflammation condition correlated positively with the inoculated virus doses. It is not surprising that virus load affects the disease progress [30]. Larger virus load is destined to bring about more severe damage in the acute phase, causing different mortality and myocardia inflammation infiltration extent.
C57BL/6 mice have been considered resistant to cardiotropic coxsackievirus B3 (CVB3)-induced myocarditis. However, establishment of viral myocarditis model in C57BL/6 mouse is still a necessity. Here, we discuss the effects of mouse initial body weight and different virus inoculation doses on the onset of acute viral myocarditis in C57BL/6 mouse.
According to initial body weight, mice were grouped into group A (3–4 weeks, 11–15 g) and group B (5–6 weeks, 18–20 g). Then, each group was divided into three subgroups inoculated with different virus doses: 1000 50% tissue culture infectious dose (TCID₅₀)/ml, 10,000 TCID₅₀/ml, and 100,000 TCID₅₀/ml. Virus existence and myocardium inflammatory infiltration conditions were observed and evaluated 7 days postinfection.
Immunohistochemistry staining showed that virus capsid protein VP1 appeared in the myocardia of virus-infected mice. Three subgroups in the lower-body-weight group (group A) came out with a higher mortality (40%–100%), while the higher-body-weight group (group B) showed a tendency for body weight decline. Histopathologically, myocardia of the survival cases in the lower-initial-body-weight group got inflammatory infiltration, while almost no inflammation appeared in the dead cases. In the higher-body-weight group, myocardium inflammatory infiltration deteriorated with the increase of virus doses and bared a relatively sound survival rate.
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ArticlesSecondary Heterotypic Versus Homotypic Infection by Coxsackie B Group Viruses: Impact on Early and Late Histopathological Lesions and Virus Genome Prominence

Abstract

Section snippets

Mice and Virus Inoculation

Virus Genome Quantitation

Discussion

Acknowledgements

J Am Coll Cardiol

Clin Immunol Immunopathol

Cell

Mol Cell Probes

Epidemiology of group B coxsackieviruses

Group B coxsackievirus infections in infants younger than three months of agea serious childhood illness [review]

Rev Infect Dis

Murine cell mediated immune response recognizes an enterovirus group specific antigen

Viral Immunol

Secondary enterovirus infection in the murine model of myocarditispathological and immunological aspects

Am J Pathol

Complete nucleotide sequence of infectious coxsackievirus B3 cDNAtwo initial 5′ uridine residues are regained during plus strand RNA synthesis

J Virol

Enteroviral infection of mice with severe combined immunodeficiency. Evidence for direct viral pathogenesis of myocardial injury

Lab Invest