Pregnant women infected with pandemic H1N1pdm2009 influenza virus displayed overproduction of peripheral blood CD69+ lymphocytes and increased levels of serum cytokines

Arturo Cérbulo-Vázquez, Ricardo Figueroa-Damián, Lourdes A Arriaga-Pizano, Edgar Hernández-Andrade, Ismael Mancilla-Herrera, Luis Angel Flores-Mejía, Gabriel Arteaga-Troncoso, Constantino López-Macías, Armando Isibasi, Javier Mancilla-Ramírez, Arturo Cérbulo-Vázquez, Ricardo Figueroa-Damián, Lourdes A Arriaga-Pizano, Edgar Hernández-Andrade, Ismael Mancilla-Herrera, Luis Angel Flores-Mejía, Gabriel Arteaga-Troncoso, Constantino López-Macías, Armando Isibasi, Javier Mancilla-Ramírez

Abstract

The first pandemic of the 21st century occurred in 2009 and was caused by the H1N1pdm influenza A virus. Severe cases of H1N1pdm infection in adults are characterized by sustained immune activation, whereas pregnant women are prone to more severe forms of influenza, with increased morbi-mortality. During the H1N1pdm09 pandemic, few studies assessed the immune status of infected pregnant women. The objective of this study was to evaluate the behavior of several immune markers in 13 H1N1pdm2009 virus-infected pregnant (PH1N1) women, in comparison to pregnant women with an influenza-like illness (ILI), healthy pregnant women (HP) and healthy non-pregnant women (HW). The blood leukocyte phenotypes and the serological cytokine and chemokine concentrations of the blood leukocytes, as measured by flow cytometry, showed that the CD69+ cell counts in the T and B-lymphocytes were significantly higher in the PH1N1 group. We found that pro-inflammatory (TNF-α, IL-1β, IL-6) and anti-inflammatory (IL-10) cytokines and some chemokines (CXCL8, CXCL10), which are typically at lower levels during pregnancy, were substantially increased in the women in the ILI group. Our findings suggest that CD69 overexpression in blood lymphocytes and elevated levels of serum cytokines might be potential markers for the discrimination of H1N1 disease from other influenza-like illnesses in pregnant women.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Higher percentage of CD69+ lymphocytes…
Figure 1. Higher percentage of CD69+ lymphocytes in H1N1pdm2009 virus-infected pregnant women.
The peripheral blood leukocytes from the HW, HP, ILI and PH1N1 women were immunostained with CD3-, CD19-, CD14- and CD69-specific antibodies and analyzed by flow cytometry. Representative data of each group are shown in the dot plots for CD69+CD3+ cells (a). The distribution of CD69 expression on CD3− (b), CD19− (c) or CD14− (d) gated cells. The Kruskal-Wallis test with Dunn’s multiple comparison post-test was performed using the GraphPad Software. The significance values were *p

Figure 2. Serum cytokine concentrations in HW,…

Figure 2. Serum cytokine concentrations in HW, HP, ILI and PH1N1 women.

Pro-inflammatory TNF-α (a),…

Figure 2. Serum cytokine concentrations in HW, HP, ILI and PH1N1 women.
Pro-inflammatory TNF-α (a), IL-1β (b) and IL-6 (c) and anti-inflammatory IL-10 (d) cytokines were quantified using a CBA system with flow cytometry. The Kruskal-Wallis test with Dunn’s multiple comparison post-test was performed using the GraphPad Software. The significance values were *p

Figure 3. Serum chemokine concentrations in HW,…

Figure 3. Serum chemokine concentrations in HW, HP, ILI and PH1N1 women.

The chemokine CXCL9/MIG…

Figure 3. Serum chemokine concentrations in HW, HP, ILI and PH1N1 women.
The chemokine CXCL9/MIG (a), CCL2/MCP-1 (b), CXCL8/IL-8 (c) and CXCL10/IP-10 (d) concentrations were quantified using a CBA system with flow cytometry. The Kruskal-Wallis test with Dunn’s multiple comparison post-test was performed using the GraphPad Software. The significance values were *p
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References
    1. Dawood FS, Jain S, Finelli L, Shaw MW, Lindstrom S, et al. (2009) Emergence of a novel swine-origin influenza A (H1N1) virus in humans. N Engl J Med 360: 2605–2615. - PubMed
    1. Perez-Padilla R, de la Rosa-Zamboni D, Ponce de Leon S, Hernandez M, Quinones-Falconi F, et al. (2009) Pneumonia and respiratory failure from swine-origin influenza A (H1N1) in Mexico. N Engl J Med 361: 680–689. - PubMed
    1. Kumar A, Zarychanski R, Pinto R, Cook DJ, Marshall J, et al. (2009) Critically ill patients with 2009 influenza A(H1N1) infection in Canada. JAMA 302: 1872–1879. - PubMed
    1. Louie JK, Acosta M, Winter K, Jean C, Gavali S, et al. (2009) Factors associated with death or hospitalization due to pandemic 2009 influenza A(H1N1) infection in California. JAMA 302: 1896–1902. - PubMed
    1. Webb SA, Pettila V, Seppelt I, Bellomo R, Bailey M, et al. (2009) Critical care services and 2009 H1N1 influenza in Australia and New Zealand. N Engl J Med 361: 1925–1934. - PubMed
Show all 59 references
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This study was supported by grants from INPer-SALUD: 2009-C02-127102 and IMSS-SALUD:2009-C02-127068. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Figure 2. Serum cytokine concentrations in HW,…
Figure 2. Serum cytokine concentrations in HW, HP, ILI and PH1N1 women.
Pro-inflammatory TNF-α (a), IL-1β (b) and IL-6 (c) and anti-inflammatory IL-10 (d) cytokines were quantified using a CBA system with flow cytometry. The Kruskal-Wallis test with Dunn’s multiple comparison post-test was performed using the GraphPad Software. The significance values were *p

Figure 3. Serum chemokine concentrations in HW,…

Figure 3. Serum chemokine concentrations in HW, HP, ILI and PH1N1 women.

The chemokine CXCL9/MIG…

Figure 3. Serum chemokine concentrations in HW, HP, ILI and PH1N1 women.
The chemokine CXCL9/MIG (a), CCL2/MCP-1 (b), CXCL8/IL-8 (c) and CXCL10/IP-10 (d) concentrations were quantified using a CBA system with flow cytometry. The Kruskal-Wallis test with Dunn’s multiple comparison post-test was performed using the GraphPad Software. The significance values were *p
Similar articles
Cited by
References
    1. Dawood FS, Jain S, Finelli L, Shaw MW, Lindstrom S, et al. (2009) Emergence of a novel swine-origin influenza A (H1N1) virus in humans. N Engl J Med 360: 2605–2615. - PubMed
    1. Perez-Padilla R, de la Rosa-Zamboni D, Ponce de Leon S, Hernandez M, Quinones-Falconi F, et al. (2009) Pneumonia and respiratory failure from swine-origin influenza A (H1N1) in Mexico. N Engl J Med 361: 680–689. - PubMed
    1. Kumar A, Zarychanski R, Pinto R, Cook DJ, Marshall J, et al. (2009) Critically ill patients with 2009 influenza A(H1N1) infection in Canada. JAMA 302: 1872–1879. - PubMed
    1. Louie JK, Acosta M, Winter K, Jean C, Gavali S, et al. (2009) Factors associated with death or hospitalization due to pandemic 2009 influenza A(H1N1) infection in California. JAMA 302: 1896–1902. - PubMed
    1. Webb SA, Pettila V, Seppelt I, Bellomo R, Bailey M, et al. (2009) Critical care services and 2009 H1N1 influenza in Australia and New Zealand. N Engl J Med 361: 1925–1934. - PubMed
Show all 59 references
Publication types
MeSH terms
Related information
Grant support
This study was supported by grants from INPer-SALUD: 2009-C02-127102 and IMSS-SALUD:2009-C02-127068. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
[x]
Cite
Copy Download .nbib
Format: AMA APA MLA NLM
Figure 3. Serum chemokine concentrations in HW,…
Figure 3. Serum chemokine concentrations in HW, HP, ILI and PH1N1 women.
The chemokine CXCL9/MIG (a), CCL2/MCP-1 (b), CXCL8/IL-8 (c) and CXCL10/IP-10 (d) concentrations were quantified using a CBA system with flow cytometry. The Kruskal-Wallis test with Dunn’s multiple comparison post-test was performed using the GraphPad Software. The significance values were *p

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