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Results: 5

1.
Fig. 3.

Fig. 3. From: Differential HIV-1 replication in neonatal and adult blood mononuclear cells is influenced at the level of HIV-1 gene expression.

FACS analysis of HIV-1 receptor (CD4) and coreceptor (CXCR4 and CCR5) expression in cord (dotted lines) vs. adult (dashed lines) blood T lymphocytes and MDM. The x axis represents the expression of the receptor/coreceptor, whereas y axis represents cell count. FACS analysis has been performed on lymphocytes and MDM from seven different donors with no significant difference in the levels of CD4, CXCR4, and CCR5.

Vasudha Sundaravaradan, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11701-11706.
2.
Fig. 2.

Fig. 2. From: Differential HIV-1 replication in neonatal and adult blood mononuclear cells is influenced at the level of HIV-1 gene expression.

Replication of HIV-1 primary isolates in cord and adult blood T lymphocytes (A) and MDM (B). T lymphocytes (1 × 106) and MDM (0.5 × 106) obtained from adult and cord blood were infected with 1 × 105 RT counts of several R5, X4, and X4/R5 HIV-1 primary isolates. Virus production was measured by RT assay in culture supernatant and three peak days (D) are shown. The results were expressed as counts per million per milliliter ± SD of triplicate experiments. These experiments have been performed in MDM and T lymphocytes from seven different cord and adult blood donors with similar results.

Vasudha Sundaravaradan, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11701-11706.
3.
Fig. 1.

Fig. 1. From: Differential HIV-1 replication in neonatal and adult blood mononuclear cells is influenced at the level of HIV-1 gene expression.

Replication of HIV-1BaL in neonatal (cord) and adult blood T lymphocytes (A) and MDM (B), and HIV-1NL4–3 in T lymphocytes (C). T lymphocytes (1 × 106) and MDM (0.5 × 106) obtained from cord and adult blood were infected with 1 × 105 RT counts of HIV-1Ba-L and HIV-1NL4–3. At different time periods, the virus production was measured in the culture supernatant by RT assay, and the two peak days are shown. The results are expressed as counts per million per milliliter ± SD. of triplicate experiments. These experiments have been performed from seven different donors with the similar statistically significant results. The P values for HIV-1Ba-L in MDM were <0.001, <0.0001 for HIV-1Ba-L in T lymphocytes, and <0.0003 for HIV-1NL4–3 in T lymphocytes (n = 7).

Vasudha Sundaravaradan, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11701-11706.
4.
Fig. 4.

Fig. 4. From: Differential HIV-1 replication in neonatal and adult blood mononuclear cells is influenced at the level of HIV-1 gene expression.

Cell proliferation ([3H]thymidine) uptake assay in adult and cord lymphocyte and MDM (A) and postentry events of HIV-1 infection in MDM (B). T lymphocytes and MDM were incubated with [3H]thymidine mock and infected with HIV-1BaL as described in Materials and Methods. As can be seen (A), there was no difference in the proliferative abilities of cord blood MDM compared with adult blood MDM. T lymphocytes from adult blood proliferated better than cord T lymphocytes. The P values for MDM are <0.01 (n = 4) and <0.05 for lymphocytes (n = 5). (B) Comparative analysis of PCR amplification products of HIV-1 postentry events, including reverse transcription, DNA synthesis, and translocation of PIC into the nucleus in adult and cord MDM. Ba, R/U5; Bb, R/U3; Bc, gag; Bd, 2LTR; Be, α-tubulin. Lanes: 1, BaL; 2, primary R5 isolate; 3, NL 4–3; 4, primary X4 isolate, 5, Mock. There was no significant difference in postentry events. In 2LTR, the X4 isolates bands are much weaker than R5 because of the block at 2LTR level. These experiments were performed in MDM from seven different cord and adult blood donors with similar results.

Vasudha Sundaravaradan, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11701-11706.
5.
Fig. 5.

Fig. 5. From: Differential HIV-1 replication in neonatal and adult blood mononuclear cells is influenced at the level of HIV-1 gene expression.

HIV-1 gene expression in cord and adult T lymphocytes (A) and MDM (B) and HIV-1 LTR transcription in MDM (C). T lymphocytes (1 × 106) and MDM (0.5 × 106) were infected with 1 × 105 RT counts of HIV-NL-LucER+ (R+E) and HIV-NL-Luc-ER (RE). Cultured cells were harvested 72 h later, and luciferase activity was assayed in cell lysate as described in Materials and Methods. The enzyme activity was normalized based on total cellular protein. The results are expressed as relative light units (RLU) ± SD of triplicate experiments. These experiments have been performed in MDM (P < 0.01) and T lymphocytes (P < 0.000001) from seven different donors of cord and adult blood. (C) Ribonuclease protection assay of luciferase mRNA transcription. Cord and adult blood MDM were infected with equal amounts of HIV-NL-Luc-ER+ (R+E) and HIV-NL-Luc-ER (RE) amphotropic viruses, and lysates were hybridized with luciferase antisense RNA probe and digested with RNase. Protected bands were analyzed on urea-PAGE. α-tubulin was used as internal control. NS, nonspecific RNA. The band in the mock is similar to nonspecific RNA. The densitometric analysis was done on RPA from five donors of cord and adult MDM (data not shown) with P values of P < 0.001.

Vasudha Sundaravaradan, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11701-11706.

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