Journal of Infectious Diseases Advance Access published February 4, 2014 1
Differences in Antibody Responses between TIV and LAIV Influenza Vaccines Correlate
cr ipt
with the Kinetics and Magnitude of Interferon Signaling in Children
Raquel G. Cao1, Nicolas M. Suarez1, Gerlinde Obermoser4, Santiago M.C. Lopez1, Emilio Flano1, Sara E. Mertz1, Randy A. Albrecht5,6, Adolfo García-Sastre5,6,7, Asuncion
us
Pascual4, Octavio Ramilo1,2,3,*
Center for Vaccines & Immunity, The Research Institute at Nationwide Children’s Hospital,
Columbus, OH
an
1
Department of Pediatrics, Division of Infectious Diseases, Nationwide Children’s Hospital
3
The Ohio State University College of Medicine, Columbus, OH
4
Baylor Institute for Immunology Research, Baylor Research Institute, Dallas, TX
5
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY
6
Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai,
pt ed
M
2
New York, NY 7
Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount
*
ce
Sinai, New York, NY
Corresponding author: Octavio Ramilo, MD, Division of Pediatric Infectious Diseases & Center
Ac
for Vaccines & Immunity, The Research Institute at Nationwide Children’s Hospital, The Ohio State University College of Medicine, 700 Children’s Drive, W4021 - Columbus, OH 43205, Phone number: 614-722-2735, Fax number: 614-722-3680, E-mail: [email protected]
© The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e‐mail: [email protected].
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
Mejias1,2,3, Hui Xu4, Huanying Qin4, Derek Blankenship4, Karolina Palucka4, Virginia
2
Abstract Background: LAIV and TIV influenza vaccines are effective for prevention of influenza
cr ipt
infection in children, but the mechanisms associated with protection are not well defined.
Methods: We analyzed the differences in B cell responses and transcriptional profiles in children 6 months to 14 years old immunized with these two vaccines.
us
post-vaccination, while TIV elicited an increased number of plasmablasts on day 7. Antibody
an
titers against the three vaccine strains (H1N1, H3N2 and B) were significantly higher in the TIV group and correlated with number of antibody-secreting cells. Both vaccines induced over expression of interferon signaling genes, but with different kinetics. TIV induced expression of
M
interferon genes on day 1 in all age groups, and LAIV on day 7 post-vaccination but only in children younger than 5 years old. Interferon-related genes over expressed in both vaccinated
pt ed
groups correlated with H3N2 antibody titers.
Conclusions: These results suggest that LAIV and TIV induced significantly different B cell responses in vaccinated children. Early induction of interferon appears important for
Ac
ce
development of antibody responses.
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
Results: LAIV elicited a significant increase in naïve, memory and transitional B cells on day 30
3
Introduction Influenza viruses cause annual epidemics and intermittent pandemics worldwide. Studies
cr ipt
have demonstrated that in addition to children with underlying conditions, influenza also causes significant morbidity and mortality healthy children (1-4). In USA influenza vaccination is recommended for all individuals 6 months and older (5).
us
trivalent inactivated influenza vaccine (TIV) and the live attenuated influenza vaccine (LAIV).
an
Both vaccines are efficacious in reducing influenza infection in children (6-9), some studies, however, suggest that LAIV has higher efficacy than TIV in young children (10-12). The immune mechanisms by which these vaccines induce immune protection are not well
M
understood. We conducted a prospective study in children immunized with TIV or LAIV to characterize the differences in: a) B cell populations by flow cytometry, b) serum antibody titers
pt ed
by hemagglutination inhibition (HAI) and viral neutralization assays (VNA), and c) whole blood transcriptional profiles to determine if early changes in expression of certain immune related
ce
genes correlated with antibody responses.
Material and Methods
Ac
Experimental Design
This was a prospective cohort of previously healthy children 6 months to 14 years of age
enrolled from October 2011 to February 2012. Forty children were initially randomized to receive one dose of LAIV (FluMist®, MedImmune) or TIV (Fluzone®, Sanofi Pasteur). Three
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
There are two major types of influenza vaccines approved for children in USA, the
4
children in the TIV cohort withdrew from the study, so the analyzed study cohort included 37 children (LAIV, n=20; TIV, n=17). Two exceptions were applied to the randomization: 4
cr ipt
children less than 2 years old and 4 children with controlled asthma received TIV following
CDC’s recommendations. One child aged 6 months at enrollment received two vaccine doses with one month interval, since it was his first influenza vaccination. In this child, all
immunologic studies were performed only after the initial vaccine dose. All other children had
us
collected. Demographic data is presented on Table 1. Exclusion criteria included chronic
an
conditions, except asthma, immunodeficiencies, and use of systemic steroids in the previous 2 weeks. Children with history of fever or respiratory symptoms within 4 weeks before enrollment
M
were also excluded.
Blood samples were collected on day 0 prior to vaccination, and days 1, 7 (range 6-8) and
pt ed
30 (range 27-33) post-vaccination to measure B-cells, antibody responses and gene expression profiles. The study used the 2011-2012 vaccines (A/California/7/2009 (H1N1)-like, A/Perth/16/2009 (H3N2)-like, B/Brisbane/60/2008-like). Adverse events were collected by the investigators at each study visit. The study was approved by the IRB at Nationwide Children’s Hospital (IRB09-00262) and Baylor Institute for Immunology Research (IRB011-221). Written
Ac
ce
informed consent was obtained from all parents.
Flow Cytometry
Peripheral whole blood samples were analyzed with the following antibody panel to
identify the B cell populations: CD45 (Pacific Orange) (Invitrogen); CD14 (Alexa Fluor 700),
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
previous influenza vaccination and precise data on the previous 2010-11 season vaccination were
5
CD20 (Pe-Cy5), IgD (FITC), CD24 (PE) (BD Pharmingen); CD19 (ECD, Beckman-Coulter); CD27 (APC-Cy7, BioLegend); CD38 (Pe-Cy7), CD138 (APC) (BD BioScience). The staining
cr ipt
protocol is described elsewhere (13). Events were collected on a LSRII instrument (BD
Biosciences, San Jose, CA) and analysis performed using FlowJo™ software (Treestar, Inc
version 9.4.11). Total cell numbers were calculated using the routine white blood cell counts
an
Hemagglutination Inhibition Assay and Virus Neutralization Assay
Serum samples were collected at two time points, day 0 and day 30 post-vaccination.
M
HAI was performed as previously described (14). The Hemmagglutination Inhibition (HI) titer was defined as the reciprocal of the highest dilution of serum that inhibits red blood cell hemagglutination. VNA was performed as previously described (15). The Viral Neutralization
pt ed
(VN) titer was defined as the reciprocal of the highest dilution of serum that neutralizes 200 PFU of influenza virus. Seroconversion was based on the following criteria: a 4-fold increase in antibody titers between the pre-vaccination and the convalescent serum samples, or an increase
18).
ce
of antibody titers from
Differences in Antibody Responses between TIV and LAIV Influenza Vaccines Correlate
cr ipt
with the Kinetics and Magnitude of Interferon Signaling in Children
Raquel G. Cao1, Nicolas M. Suarez1, Gerlinde Obermoser4, Santiago M.C. Lopez1, Emilio Flano1, Sara E. Mertz1, Randy A. Albrecht5,6, Adolfo García-Sastre5,6,7, Asuncion
us
Pascual4, Octavio Ramilo1,2,3,*
Center for Vaccines & Immunity, The Research Institute at Nationwide Children’s Hospital,
Columbus, OH
an
1
Department of Pediatrics, Division of Infectious Diseases, Nationwide Children’s Hospital
3
The Ohio State University College of Medicine, Columbus, OH
4
Baylor Institute for Immunology Research, Baylor Research Institute, Dallas, TX
5
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY
6
Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai,
pt ed
M
2
New York, NY 7
Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount
*
ce
Sinai, New York, NY
Corresponding author: Octavio Ramilo, MD, Division of Pediatric Infectious Diseases & Center
Ac
for Vaccines & Immunity, The Research Institute at Nationwide Children’s Hospital, The Ohio State University College of Medicine, 700 Children’s Drive, W4021 - Columbus, OH 43205, Phone number: 614-722-2735, Fax number: 614-722-3680, E-mail: [email protected]
© The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e‐mail: [email protected].
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
Mejias1,2,3, Hui Xu4, Huanying Qin4, Derek Blankenship4, Karolina Palucka4, Virginia
2
Abstract Background: LAIV and TIV influenza vaccines are effective for prevention of influenza
cr ipt
infection in children, but the mechanisms associated with protection are not well defined.
Methods: We analyzed the differences in B cell responses and transcriptional profiles in children 6 months to 14 years old immunized with these two vaccines.
us
post-vaccination, while TIV elicited an increased number of plasmablasts on day 7. Antibody
an
titers against the three vaccine strains (H1N1, H3N2 and B) were significantly higher in the TIV group and correlated with number of antibody-secreting cells. Both vaccines induced over expression of interferon signaling genes, but with different kinetics. TIV induced expression of
M
interferon genes on day 1 in all age groups, and LAIV on day 7 post-vaccination but only in children younger than 5 years old. Interferon-related genes over expressed in both vaccinated
pt ed
groups correlated with H3N2 antibody titers.
Conclusions: These results suggest that LAIV and TIV induced significantly different B cell responses in vaccinated children. Early induction of interferon appears important for
Ac
ce
development of antibody responses.
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
Results: LAIV elicited a significant increase in naïve, memory and transitional B cells on day 30
3
Introduction Influenza viruses cause annual epidemics and intermittent pandemics worldwide. Studies
cr ipt
have demonstrated that in addition to children with underlying conditions, influenza also causes significant morbidity and mortality healthy children (1-4). In USA influenza vaccination is recommended for all individuals 6 months and older (5).
us
trivalent inactivated influenza vaccine (TIV) and the live attenuated influenza vaccine (LAIV).
an
Both vaccines are efficacious in reducing influenza infection in children (6-9), some studies, however, suggest that LAIV has higher efficacy than TIV in young children (10-12). The immune mechanisms by which these vaccines induce immune protection are not well
M
understood. We conducted a prospective study in children immunized with TIV or LAIV to characterize the differences in: a) B cell populations by flow cytometry, b) serum antibody titers
pt ed
by hemagglutination inhibition (HAI) and viral neutralization assays (VNA), and c) whole blood transcriptional profiles to determine if early changes in expression of certain immune related
ce
genes correlated with antibody responses.
Material and Methods
Ac
Experimental Design
This was a prospective cohort of previously healthy children 6 months to 14 years of age
enrolled from October 2011 to February 2012. Forty children were initially randomized to receive one dose of LAIV (FluMist®, MedImmune) or TIV (Fluzone®, Sanofi Pasteur). Three
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
There are two major types of influenza vaccines approved for children in USA, the
4
children in the TIV cohort withdrew from the study, so the analyzed study cohort included 37 children (LAIV, n=20; TIV, n=17). Two exceptions were applied to the randomization: 4
cr ipt
children less than 2 years old and 4 children with controlled asthma received TIV following
CDC’s recommendations. One child aged 6 months at enrollment received two vaccine doses with one month interval, since it was his first influenza vaccination. In this child, all
immunologic studies were performed only after the initial vaccine dose. All other children had
us
collected. Demographic data is presented on Table 1. Exclusion criteria included chronic
an
conditions, except asthma, immunodeficiencies, and use of systemic steroids in the previous 2 weeks. Children with history of fever or respiratory symptoms within 4 weeks before enrollment
M
were also excluded.
Blood samples were collected on day 0 prior to vaccination, and days 1, 7 (range 6-8) and
pt ed
30 (range 27-33) post-vaccination to measure B-cells, antibody responses and gene expression profiles. The study used the 2011-2012 vaccines (A/California/7/2009 (H1N1)-like, A/Perth/16/2009 (H3N2)-like, B/Brisbane/60/2008-like). Adverse events were collected by the investigators at each study visit. The study was approved by the IRB at Nationwide Children’s Hospital (IRB09-00262) and Baylor Institute for Immunology Research (IRB011-221). Written
Ac
ce
informed consent was obtained from all parents.
Flow Cytometry
Peripheral whole blood samples were analyzed with the following antibody panel to
identify the B cell populations: CD45 (Pacific Orange) (Invitrogen); CD14 (Alexa Fluor 700),
Downloaded from http://jid.oxfordjournals.org/ at Belgorod State University on February 10, 2014
previous influenza vaccination and precise data on the previous 2010-11 season vaccination were
5
CD20 (Pe-Cy5), IgD (FITC), CD24 (PE) (BD Pharmingen); CD19 (ECD, Beckman-Coulter); CD27 (APC-Cy7, BioLegend); CD38 (Pe-Cy7), CD138 (APC) (BD BioScience). The staining
cr ipt
protocol is described elsewhere (13). Events were collected on a LSRII instrument (BD
Biosciences, San Jose, CA) and analysis performed using FlowJo™ software (Treestar, Inc
version 9.4.11). Total cell numbers were calculated using the routine white blood cell counts
an
Hemagglutination Inhibition Assay and Virus Neutralization Assay
Serum samples were collected at two time points, day 0 and day 30 post-vaccination.
M
HAI was performed as previously described (14). The Hemmagglutination Inhibition (HI) titer was defined as the reciprocal of the highest dilution of serum that inhibits red blood cell hemagglutination. VNA was performed as previously described (15). The Viral Neutralization
pt ed
(VN) titer was defined as the reciprocal of the highest dilution of serum that neutralizes 200 PFU of influenza virus. Seroconversion was based on the following criteria: a 4-fold increase in antibody titers between the pre-vaccination and the convalescent serum samples, or an increase
18).
ce
of antibody titers from
