12 research outputs found
Viral spread of wt VACV in infected animals.
<p>Groups of 3 C57BL/6 mice were infected intranasally with 10<sup>6</sup> pfu of wt VACV. Animals remained untreated (A) or were treated one day post-infection with 10<sup>7</sup> pfu of v50ĪB13RMĪ³ (B). Replication of v50ĪB13RMĪ³ in the tissues from the treated group was examined by X-gal staining (C). Error bars indicate the standard error of the mean. Data represent a pool of two independent experiments using 3 mice per group. </p
Measurement of IFN-Ī³ in the serum following treatment.
<p>Groups of 5 C57BL/6 mice were mock-infected or infected IN with 10<sup>6</sup> pfu (~100 LD<sub>50s</sub>) of wt VACV. One day post infection mice were either mock-treated or treated with 10<sup>7</sup> pfu of v50ĪB13R or v50ĪB13RMĪ³. After 24 hours, serum was harvested and an ELISA was performed to measure IFN-Ī³. The horizontal line indicates the average and error bars indicate the standard error of the mean. Comparisons were done using the Mann-Whitney test. One asterisk (*) represents p<0.008 in comparison to the wt/v50ĪB13RMĪ³ group. Two asterisks (**) represents p<0.016 in comparison to the wt/v50ĪB13RMĪ³ group. </p
Protection of wt VACV infected animals by post-exposure vaccination with v50ĪB13RMĪ³ at one, two and three days post-infection.
<p>Groups of 5 to 10 C57BL/6 mice were infected intranasally with 10<sup>6</sup> pfu (~100 LD<sub>50s</sub>) of wt VACV. Animals were treated with 10<sup>7</sup> pfu of v50ĪB13RMĪ³ at one (ā²), two (ā) or three (ā¦) days post-infection. One group was infected with 10<sup>7</sup> pfu (~1000 LD<sub>50s</sub>) of wt VACV and treated with 10<sup>7</sup> pfu of v50ĪB13RMĪ³ at one day post-infection (ā). Controls animals were infected with wt VACV and then mock-treated (ā ), or animals were mock-infected and mock-treated (ā). (A) Comparison of survival curves was done using the log-rank test. (B) Recovery of wt VACV infected animals by post-exposure vaccination with v50ĪB13RMĪ³ at one, two and three days post-infection. The graph indicates the relative sickness of each group of animals during the course of the infection. Lines ending prematurely indicate death of all the animals from the group. A value of 0 indicates that all the animals from that group were healthy.</p
Post-exposure protection of animals after intranasal infection with a lethal dose of wt VACV and subsequent treatment with different VACV mutants.
<p>(A) Weight loss. Groups of 5 to 10 four-week-old C57BL/6 mice were infected intranasally with 10<sup>6</sup> pfu (~100 LD<sub>50s</sub>) of wt VACV and treated 1 dpi with 10<sup>7</sup> pfu of VACVE3LĪ7C (ā), VACVE3LĪ26C (ā), VACVĪE3L (ā¦), VACVĪE3L::ATVeIF2Ī± (ā”), v50ĪB13RMĪ³ (ā²) and v50ĪB13RMIL-18 (ā). There were two groups of control animals, one group was infected with wt VACV alone and was mock-treated (ā ), the second group of animals was mock-infected and mock-treated (ā). Each mouse was weighed at the indicated times. Average percentage of initial weight of the animals infected with each virus is plotted versus time (days post-infection). Lines end at the death of one animal. Error bars indicate the standard error of the mean. (B) Survival curve. The data represent a pool of two independent experiments using group sizes of 5 mice. (C) v50ĪB13RMĪ³ dose response. Groups of 10 four-week-old C57BL/6 mice were infected intranasally with 10<sup>6</sup> pfu of wt VACV (~100 LD<sub>50s</sub>) and treated one day post-infection with doses of 10<sup>2</sup>, 10<sup>3</sup>, 10<sup>4</sup>, 10<sup>5</sup>, 10<sup>6</sup>, 10<sup>7</sup> and 5 x 10<sup>8</sup> pfu of v50ĪB13RMĪ³ (ā²).</p
Histopathologic comparison and infection progression in the nasal cavity section.
<p>Mice were infected IN with 10<sup>6</sup> pfu (~100 LD<sub>50s</sub>) of wt VACV and treated one day post-infection with 10<sup>7</sup> pfu of v50ĪB13RMĪ³. Mice were sacrificed at 7-8 days post-infection. All representative sections were stained with polyclonal antibodies against VACV. S=septum, T=maxilloturbinate, M=meatus (air passage), i=incisor. (A) Maxilloturbinate section, 2 mm depth, 100X magnification (B) Whole section, 5-6 mm depth. 10X magnification. Lines indicate 200 Ī¼m length.</p
Post-exposure protection of wt VACV infected animals treated with v50ĪB13RMĪ³ using different routes of treatment.
<p>Groups of 12 to 15 C57BL/6 mice were infected intranasally with 10<sup>6</sup> pfu of wt VACV. Animals were treated one day post-infection with 10<sup>7</sup> pfu of v50ĪB13RMĪ³ intranasally, IN (ā²), intranasally using the other nostril, INON (ā), intramuscularly, IM (ā) or via scarification, SCA (<b>ā</b>). One group of animals was infected with wt VACV and then mock-treated (ā ), another group was mock-infected and mock-treated (ā). Comparison of survival curves was done using the log-rank test.</p
Protection of wt ECTV infected animals by post-exposure vaccination with v50ĪB13RMĪ³ at one day post-infection.
<p>Groups of 30 C57BL/6 mice were infected intranasally with 5 x 10<sup>3</sup> pfu (~50 LD<sub>50s</sub>) of wt ECTV. Animals were treated by FP route with saline (ā ), 3 x 10<sup>7</sup> pfu of v50ĪB13RMĪ³ (ā²) or v50ĪB13R (ā) at one day post-infection. One group of animals was mock infected and mock-treated (ā). The data were pooled from two independent experiments using group sizes of 10 and 20 mice. <i>P</i> values (log-rank test) show the significance of difference with respect to ECTV/Mock. The boxed <i>P</i> value shows the significance of the difference between v50ĪB13RMĪ³ and v50ĪB13R.</p
Figure 5
<p>Augmented CD4<sup>+</sup> HIV-1 immune responses to Gag peptide pools in exposed uninfected neonatal cord blood upon the removal of CD4<sup>+</sup>CD25<sup>+</sup> Treg cells. A) IL-2 production by undepleted whole cord blood and peripheral blood mononuclear cells (MNCs) derived CD4<sup>+</sup> T-cells (open white symbols) and CD25-depleted MNCs derived CD4<sup>+</sup> T-cells (closed black symbols) is depicted. B) Flow cytometry plots from an exposed uninfected neonate (Patient 63) representing HIV-1 Gag induced IL-2 production in undepleted whole CBMC derived CD4<sup>+</sup> T-cells and CD25-depleted CBMC derived CD4<sup>+</sup> T-cells.</p
Figure 1
<p>CD4<sup>+</sup> and CD8<sup>+</sup> T cell immune responses were measured by cytokine flow cytometry. A) Gating strategy for the identification of polyfunctional IFN-gamma/TNF-alpha CD8<sup>+</sup> T cell responses. B) Shown are representative data for the unstimulated and HIV-gag-specific response from subject PB-INF-4 after an 18 h <i>in vitro</i> stimulation.</p
Figure 3
<p>Polyfunctional CD8<sup>+</sup> T cell immune responses to the HIV-1 Gag peptide pool were detected by cytokine flow cytometry. Responses were measured in the cord blood of unexposed neonates (CB-UNEX; nā=ā4), HIV-1-exposed uninfected neonates (CB-EU; nā=ā6), and in the peripheral blood of HIV-1-exposed-uninfected infants (PB-EU 7 mo; nā=ā9) and young children (PB-EU 20 mo; nā=ā7), and in HIV-1-infected infants (PB-INF 7 mo; nā=ā5) and young children (PB-INF 25 mo; nā=ā5). Each group is represented by a different symbol.</p