Comparison of Th-cell immunity against human bocavirus and parvovirus B19: proliferation and cytokine responses are similar in magnitude but more closely interrelated with human bocavirus.

Human parvovirus B19 (B19) has been, for decades, the only parvovirus known to be pathogenic in humans. Another pathogenic human parvovirus, human bocavirus (HBoV), was recently identified in respiratory samples from children with acute lower respiratory tract symptoms. Both B19 and HBoV are transmitted by the respiratory route. The vast majority of adults are IgG seropositive for HBoV, whereas the HBoV-specific Th-cell immunity has not much been studied. The aim of this study was to increase our knowledge on HBoV-specific Th-cell immunity by examining HBoV-specific T-cell proliferation, Interferon-gamma (IFN-γ), IL-10 and IL-13 responses in 36 asymptomatic adults. Recombinant HBoV VP2 virus-like particles (VLP) were used as antigen. HBoV-specific responses were compared with those elicited by B19 VP2 VLP. Proliferation, IFN-γ and IL-10 responses with HBoV and B19 antigens among B19-seropositive subjects were statistically similar in magnitude, but the cytokine and proliferation responses were much more closely correlated in HBoV than in B19. Therefore, at the collective level, B19-specific Th-cell immunity appears to be more divergent than the HBoV-specific one.

Parvovirus B19 VP2-proteins produced in Saccharomyces cerevisiae: comparison with VP2-particles produced by baculovirus-derived vectors.

The capsids of human parvovirus B19 consist of two structural proteins, the minor-capsid protein VP1 and the major-capsid protein VP2. The latter which constitutes for 95% of the capsid are able to form virus-like particles (VLPs) in yeast without the presence of VP1-proteins. VP2-proteins produced in Saccharomyces cerevisiae have the capacity to form VLPs in the absence of VP1-proteins. These yeast-derived VLPs resemble native virus or recombinant VP2-VLPs produced by baculovirus systems in respect of size, molecular weight and of antigenicity as shown by antigen-capture ELISA and T-cell proliferation tests. Regarding costs, yield and ease of handling particle production in yeast represents an alternative to the recombinant baculovirus expression system which is so far the source for VP2-VLPs of human parvovirus B19.

T helper cell-mediated interferon-gamma expression after human parvovirus B19 infection: persisting VP2-specific and transient VP1u-specific activity.

Human parvovirus B19 is a small non-enveloped DNA virus with an icosahedral capsid consisting of proteins of only two species, the major protein VP2 and the minor protein VP1. VP2 is contained within VP1, which has an additional unique portion (VP1u) of 227 amino acids. We determined the ability of eukaryotically expressed parvovirus B19 virus-like particles consisting of VP1 and VP2 in the ratio recommended for vaccine use, or of VP2 alone, to stimulate, in an HLA class II restricted manner, peripheral blood mononuclear cells (PBMC) to proliferate and to secrete interferon gamma (IFN-gamma) and interleukin (IL)-10 cytokines among recently and remotely B19 infected subjects. PBMC reactivity with VP1u was determined specifically with a prokaryotically expressed VP1u antigen. In general, B19-specific IFN-gamma responses were stronger than IL-10 responses in both recent and remote infection; however, IL-10 responses were readily detectable among both groups, with the exception of patients with relapsed or persisting symptoms who showed strikingly low IL-10 responses. Whereas VP1u-specific IFN-gamma responses were very strong among the recently infected subjects, the VP1u-specific IFN-gamma and IL-10 responses were virtually absent among the remotely infected subjects. The disappearance of VP1u-specific IFN-gamma expression is surprising, as B-cell immunity against VP1u is well maintained.

T helper cell-mediated in vitro responses of recently and remotely infected subjects to a candidate recombinant vaccine for human parvovirus b19.

T cell proliferation to human parvovirus B19 antigen was measured in 6 patients with recent B19 infection (1 with pneumonia and pleuritis), 1 patient with symptoms persisting >180 days after onset, 18 nonsymptomatic subjects with remote B19 immunity, and 12 B19-seronegative control subjects. Recombinantly expressed virus-like particles (VP1/2 capsids), a candidate B19 vaccine, were used as antigen. Virus-specific T helper cell proliferation was detectable in all the recently infected patients and in most (17/18) of the remotely infected subjects but not in the seronegative control subjects. The B19-specific T cell responses, in general, were most vigorous among the recently infected patients. However, such strong B19-specific proliferation was not confined within the acute phase, as 28% (5/18) of the remotely infected healthy individuals had B19-specific reactivity persisting at acute-phase levels, apparently for years or decades. These data indicate that B cells recognizing the VP1/2 capsids receive class II-restricted help from CD4(+) T lymphocytes.

Primary and secondary infections by human parvovirus B19 following bone marrow transplantation: characterization by PCR and B-cell molecular immunology.

Due to the preparative regimen necessary, bone marrow transplantation (BMT) consistently results in severe immunodeficiency, often associated with anaemia, leukopenia and thrombocytopenia. Parvovirus B19 replicates in red blood cell precursors in the bone marrow and causes erythema infectiosum ('fifth disease'), anaemia, arthritis and foetal death. We assessed the significance of B19 infections as a cause of post-BMT complications. Over 900 serial serum samples from 201 allogeneic bone marrow recipients were studied by polymerase chain reaction (PCR) and by modern serodiagnostic methods. During the first 6 months after transplantation all BMT recipients remained B19 PCR-negative. Antibody screening for B19 infections was performed up to 36 months post-transplantation. Three cases of acute B19 infection were diagnosed during the second year post-BMT. To characterize the adoptively transferred immune system we measured subclasses and avidity of anti-VP1 IgG and epitope-type specificity (ETS) of anti-VP2 IgG, which allowed functional differentiation of primary and secondary B-cell responses long after BMT. The profile of the immune response was that of a primary infection in 1 and of reinfection in 2 of the 3 acute cases. Both types were clinically mild. Infection by human parvovirus B19 is not a frequent cause of post-BMT cytopenias. The findings with the new B19 antibody markers support the concept that the donated marrow determines the type of antiviral B-cell responses.

IgG subclass response to human parvovirus B19 infection.

BACKGROUND: IgG antibodies are essential to immunity against human parvovirus B19 and can neutralize infection both in bone marrow cell cultures infected in vitro and in chronically infected immunosuppressed individuals. OBJECTIVES: To assess the levels and response kinetics of IgG subclasses towards individual structural proteins of human parvovirus B19. STUDY DESIGN: Subclasses of IgG for capsid proteins VP1 or VP2 were quantified by EIA using monoclonal antibodies in 30 acutely infected and 30 convalescent patients, as well as in 32 remotely infected and 20 non-infected controls. RESULTS: In all groups of seropositive individuals the predominant subclass for either structural protein was IgG1. Subclass IgG3 was associated with acute infection. By contrast, IgG4 appeared months after infection, and occurred specifically towards VP1. The ratio of VP1-specific subclasses IgG3 and IgG4 provided a diagnostic test for recent infection with a specificity of 98% and a sensitivity of 97%. CONCLUSIONS: Comparative measurement of VP1-specific IgG3 and IgG4 is useful in diagnosis. The IgG4 results point to long-term expression of immunologically active VP1 and to T-cell help of T(h)2 type for B-cells recognizing VP1.

Contrast sensitivity as a function of spatial frequency, viewing distance and eccentricity with and without spatial noise.

Using computer graphics and a two-alternative forced-choice method we measured threshold contrast as a function of viewing distance, spatial frequency, and eccentricity for gratings with and without added, white two-dimensional spatial noise. Our experiments showed that in spatial noise contrast sensitivity was independent of viewing distance as long as contrast sensitivity was lower with noise than without. With increasing spatial frequency (f) the grating area (A) was reduced in order to keep the relative grating size (Af2) constant. At all spatial frequencies the test gratings thus had the same amount of detail and contour. Noise spectral density was reduced in direct proportion to grating area in order to keep the physical signal-to-noise ratio constant. An increase in spatial frequency was thus accompanied with reductions in grating area and noise spectral density similar to those produced by a corresponding increase in viewing distance. In agreement, contrast detection in spatial noise was found to be independent of spatial frequency as long as contrast sensitivity was lower with noise than without. The effect of increasing eccentricity on visual performance can be compensated for by reducing the viewing distance (M-scaling). Hence, without M-scaling the effect of increasing eccentricity is similar to that of increasing viewing distance. In agreement, we found that contrast sensitivity in spatial noise was independent of eccentricity as long as contrast sensitivity was lower with noise than without.

Critical flicker frequency to red targets as a function of luminance and flux across the human visual field.

When the number of cells (cones at eccentricities 0-10 deg and ganglion cells above 10 deg) stimulated at various retinal locations was kept constant by enlarging the stimulus area with increasing eccentricity in the temporal visual field (M-scaling), CFF to red stimuli with dark surround increased as a single function of photopic luminous flux, collected by ganglion-cell receptive-field centres and calculated by multiplying Ricco's area with retinal illuminance at each eccentricity studied. The increase of CFF with the logarithm of photopic flux could be best explained by the Collins logarithmic law, the Kelly square-root law was almost equally good and the Ferry-Porter law was poorest. Adopting the general formulation of Corwin and Dunlap (Vision Research, 27, 2119-2123, 1987) the exponent of CFF is 0, 0.5, and 1 for the Collins, Kelly and Ferry-Porter laws, respectively. The exponent that best explained our results was found to be 0-0.3.