Development and evaluation of an M2-293FT cell-based flow cytometric assay for quantification of antibody response to native form of matrix protein 2 of influenza A viruses

2011 ◽  
Vol 369 (1-2) ◽  
pp. 115-124 ◽  
Author(s):  
Weimin Zhong ◽  
Ju He ◽  
Xiaoling Tang ◽  
Feng Liu ◽  
Xiuhua Lu ◽  
...  
Keyword(s):  
Antibody Response ◽  
Influenza A ◽  
Matrix Protein ◽  
Flow Cytometric Assay ◽  
Influenza A Viruses ◽  
Native Form ◽  
Flow Cytometric ◽  
293Ft Cell

scholarly journals The impact of vaccination on the breadth and magnitude of the antibody response to influenza A viruses in HIV-infected individuals

AIDS ◽  
2015 ◽  
Vol 29 (14) ◽  
pp. 1803-1810 ◽  
Author(s):  
Ines Kohler ◽  
Roger Kouyos ◽  
Matteo Bianchi ◽  
Christina Grube ◽  
Arkadiusz Wyrzucki ◽  
...  
Keyword(s):  
Antibody Response ◽  
Influenza A ◽  
Influenza A Viruses ◽  
The Impact

scholarly journals Cooperation between the Hemagglutinin of Avian Viruses and the Matrix Protein of Human Influenza A Viruses

2002 ◽  
Vol 76 (4) ◽  
pp. 1781-1786 ◽  
Author(s):  
Christoph Scholtissek ◽  
Jürgen Stech ◽  
Scott Krauss ◽  
Robert G. Webster
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Matrix Protein ◽  
Gene Products ◽  
Human Influenza ◽  
Influenza A Viruses ◽  
M Gene ◽  
Human Virus ◽  
Ha Gene ◽  
M Proteins

ABSTRACT To analyze the compatibility of avian influenza A virus hemagglutinins (HAs) and human influenza A virus matrix (M) proteins M1 and M2, we doubly infected Madin-Darby canine kidney cells with amantadine (1-aminoadamantane hydrochloride)-resistant human viruses and amantadine-sensitive avian strains. By using antisera against the human virus HAs and amantadine, we selected reassortants containing the human virus M gene and the avian virus HA gene. In our system, high virus yields and large, well-defined plaques indicated that the avian HAs and the human M gene products could cooperate effectively; low virus yields and small, turbid plaques indicated that cooperation was poor. The M gene products are among the primary components that determine the species specificities of influenza A viruses. Therefore, our system also indicated whether the avian HA genes effectively reassorted into the genome and replaced the HA gene of the prevailing human influenza A viruses. Most of the avian HAs that we tested efficiently cooperated with the M gene products of the early human A/PR/8/34 (H1N1) virus; however, the avian HAs did not effectively cooperate with the most recently isolated human virus that we tested, A/Nanchang/933/95 (H3N2). Cooperation between the avian HAs and the M proteins of the human A/Singapore/57 (H2N2) virus was moderate. These results suggest that the currently prevailing human influenza A viruses might have lost their ability to undergo antigenic shift and therefore are unable to form new pandemic viruses that contain an avian HA, a finding that is of great interest for pandemic planning.

scholarly journals Enhanced Influenza Virus-Like Particle Vaccines Containing the Extracellular Domain of Matrix Protein 2 and a Toll-Like Receptor Ligand

2012 ◽  
Vol 19 (8) ◽  
pp. 1119-1125 ◽  
Author(s):  
Bao-Zhong Wang ◽  
Harvinder S. Gill ◽  
Sang-Moo Kang ◽  
Li Wang ◽  
Ying-Chun Wang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Fusion Protein ◽  
Influenza A ◽  
Matrix Protein ◽  
Protective Efficacy ◽  
Extracellular Domain ◽  
Toll Like Receptor ◽  
Influenza A Viruses ◽  
Live Virus ◽  
Content Type

ABSTRACTThe extracellular domain of matrix protein 2 (M2e) is conserved among influenza A viruses. The goal of this project is to develop enhanced influenza vaccines with broad protective efficacy using the M2e antigen. We designed a membrane-anchored fusion protein by replacing the hyperimmunogenic region ofSalmonella entericaserovar Typhimurium flagellin (FliC) with four repeats of M2e (4.M2e-tFliC) and fusing it to a membrane anchor from influenza virus hemagglutinin (HA). The fusion protein was incorporated into influenza virus M1-based virus-like particles (VLPs). These VLPs retained Toll-like receptor 5 (TLR5) agonist activity comparable to that of soluble FliC. Mice immunized with the VLPs by either intramuscular or intranasal immunization showed high levels of systemic M2-specific antibody responses compared to the responses to soluble 4.M2e protein. High mucosal antibody titers were also induced in intranasally immunized mice. All intranasally immunized mice survived lethal challenges with live virus, while intramuscularly immunized mice showed only partial protection, revealing better protection by the intranasal route. These results indicate that a combination of M2e antigens and TLR ligand adjuvants in VLPs has potential for development of a broadly protective influenza A virus vaccine.

scholarly journals Serum Antibody Response to Matrix Protein 2 Following Natural Infection With 2009 Pandemic Influenza A(H1N1) Virus in Humans

2013 ◽  
Vol 209 (7) ◽  
pp. 986-994 ◽  
Author(s):  
Weimin Zhong ◽  
Carrie Reed ◽  
Patrick J. Blair ◽  
Jacqueline M. Katz ◽  
Kathy Hancock ◽  
...  
Keyword(s):  
Antibody Response ◽  
Pandemic Influenza ◽  
Influenza A ◽  
Matrix Protein ◽  
H1n1 Virus ◽  
Natural Infection ◽  
Serum Antibody ◽  
Serum Antibody Response ◽  
Influenza A H1n1

scholarly journals Potentiation of Recombinant NP and M1-Induced Cellular Immune Responses and Protection by Physical Radiofrequency Adjuvant

Vaccines ◽  
2021 ◽  
Vol 9 (12) ◽  
pp. 1382
Author(s):  
Yibo Li ◽  
Zhuofan Li ◽  
Yiwen Zhao ◽  
Xinyuan Chen
Keyword(s):  
Immune Responses ◽  
Influenza A ◽  
Matrix Protein ◽  
Body Weight Loss ◽  
Antibody Responses ◽  
Significant Protection ◽  
Influenza A Viruses ◽  
Humoral Immune ◽  
Reduced Body ◽  
Ctl Responses

Nucleoprotein (NP) and matrix protein 1 (M1) are highly conserved among influenza A viruses and have been attractive targets to develop vaccines to elicit cross-reactive cytotoxic T lymphocytes (CTLs). Yet, external antigens are often presented on major histocompatibility complex class II molecules and elicit humoral immune responses. In this study, we present a physical radiofrequency adjuvant (RFA) to assist recombinant NP and M1 to elicit potent CTL responses. We found recombinant NP/M1 immunization in the presence of RFA could elicit potent anti-NP CTLs and confer significant protection against homologous viral challenges, while NP/M1 immunization alone failed to elicit significant CTL responses or confer significant protection. Interestingly, RFA failed to elicit potent anti-M1 CTL responses or anti-NP or anti-M1 antibody responses. Different from RFA, AddaVax adjuvant was found to significantly increase NP-specific antibody responses but not CTLs. NP/M1 immunization in the presence of RFA or AddaVax similarly reduced body weight loss, while only the former significantly increased the survival. We further found NP/M1 immunization in the presence of RFA did not significantly increase serum IL-6 release (a systemic inflammatory mediator) and rather reduced serum IL-6 release after boost immunization. NP/M1 immunization in the presence of RFA did not induce significant local reactions or increase body temperature of mice. The high potency and safety strongly support further development of RFA-based recombinant NP/M1 vaccine to elicit cross-protective immunity.

UBIQUITOUS REASSORTMENTS IN INFLUENZA A VIRUSES

2008 ◽  
Vol 06 (05) ◽  
pp. 981-999 ◽  
Author(s):  
XIU-FENG WAN ◽  
MUFIT OZDEN ◽  
GUOHUI LIN
Keyword(s):  
Influenza A ◽  
Matrix Protein ◽  
Minimum Spanning Tree ◽  
Migratory Birds ◽  
Rna Virus ◽  
Influenza Pandemic ◽  
Influenza Viruses ◽  
Influenza A Viruses ◽  
H5n1 Avian Influenza Virus ◽  
Pandemic Strains

The influenza A virus is a negative-stranded RNA virus composed of eight segmented RNA molecules, including polymerases (PB2, PB1, PA), hemagglutinin (HA), nucleoprotein (NP), neuraminidase (NA), matrix protein (MP), and nonstructure gene (NS). The influenza A viruses are notorious for rapid mutations, frequent reassortments, and possible recombinations. Among these evolutionary events, reassortments refer to exchanges of discrete RNA segments between co-infected influenza viruses, and they have facilitated the generation of pandemic and epidemic strains. Thus, identification of reassortments will be critical for pandemic and epidemic prevention and control. This paper presents a reassortment identification method based on distance measurement using complete composition vector (CCV) and segment clustering using a minimum spanning tree (MST) algorithm. By applying this method, we identified 34 potential reassortment clusters among 2,641 PB2 segments of influenza A viruses. Among the 83 serotypes tested, at least 56 (67.46%) exchanged their fragments with another serotype of influenza A viruses. These identified reassortments involve 1,957 H2N1 and 1,968 H3N2 influenza pandemic strains as well as H5N1 avian influenza virus isolates, which have generated the potential for a future pandemic threat. More frequent reassortments were found to occur in wild birds, especially migratory birds. This MST clustering program is written in Java and will be available upon request.

scholarly journals Virus-neutralizing antibody response of mice to consecutive infection with human and avian influenza A viruses

2015 ◽  
Vol 59 (02) ◽  
pp. 166-173
Author(s):  
J. JANULÍKOVÁ ◽  
A. STROPKOVSKÁ ◽  
Z. BOBIŠOVÁ ◽  
I. KOŠÍK ◽  
V. MUCHA ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Antibody Response ◽  
Influenza A ◽  
Neutralizing Antibody ◽  
Influenza A Viruses

scholarly journals The substantia nigra is a major target for neurovirulent influenza A virus.

1995 ◽  
Vol 181 (6) ◽  
pp. 2161-2169 ◽  
Author(s):  
M Takahashi ◽  
T Yamada ◽  
S Nakajima ◽  
K Nakajima ◽  
T Yamamoto ◽  
...  
Keyword(s):  
Substantia Nigra ◽  
Influenza A Virus ◽  
Influenza A ◽  
Matrix Protein ◽  
Gene Segment ◽  
Positive Staining ◽  
Influenza A Viruses ◽  
Postencephalitic Parkinsonism ◽  
Causative Agents ◽  
Immunohistochemical Studies

Clinical and immunohistochemical studies were done for 3-39 d on mice after intracerebral inoculation with the neurovirulent A/WSN/33 (H1N1; WSN) strain of influenza A virus, the nonneurovirulent A/Aichi/2/68 (H3N2; Aichi) strain, and two reassortant viruses between them. The virus strains with the WSN gene segment coding for neuraminidase induced meningoencephalitis in mice. The mice inoculated with the R96 strain, which has only the neuraminidase gene from the WSN strain, had mild symptoms and weak positive immunostaining to the anti-WSN antibody in meningeal regions. Both the WSN and R404BP strains, which contain the WSN gene segments coding for neuraminidase and matrix protein, were clearly neurovirulent both clinically and pathologically. On day 3 after inoculation with either of these two strains, WSN antigen was detected in meningeal and ependymal areas, neurons of circumventricular regions, the cerebral and cerebellar cortices, the substantia nigra zona compacta, and the ventral tegmental area. On day 7, meningeal reactions and neuronal staining were still seen, and advanced accumulation of the viral antigen was evident in the substantia nigra zona compacta and hippocampus. Double immunostaining demonstrated that the WSN antigen was only seen in neurons and not in microglia or reactive astrocytes. Immunostaining for the lectin maackia amurensis agglutinin, which recognizes the Neu5Ac alpha 2,3 Gal sequence, which serves as a binding site for influenza A virus on target cell membranes, showed that positive staining was localized in the ventral substantia nigra and hippocampus. These results suggest that neurovirulent influenza A viruses could be one of the causative agents for postencephalitic parkinsonism.

scholarly journals Production and Characterization of a Human Recombinant Monoclonal Fab Fragment Specific for Influenza A Viruses

2003 ◽  
Vol 10 (4) ◽  
pp. 680-685 ◽  
Author(s):  
Alessandra Desogus ◽  
Roberto Burioni ◽  
Angela Ingianni ◽  
Francesca Bugli ◽  
Raffaello Pompei ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Influenza A ◽  
Matrix Protein ◽  
Viral Infections ◽  
Mdck Cells ◽  
Influenza B Virus ◽  
Influenza B ◽  
Influenza A Viruses ◽  
Fab Fragment ◽  
Phage Display Technique

ABSTRACT A human recombinant monoclonal Fab fragment that specifically recognizes all the influenza A virus strains tested was produced in transformed Escherichia coli using the phage display technique. No strain of influenza B virus reacted with it. It was purified after four cycles of panning and by a single passage through an immunoaffinity column. About 1 mg of pure monoclonal antibody was obtained from 1 liter of culture medium in 3 working days. The Fab fragment reacted with a viral 27-kDa protein, which could reasonably be a matrix protein. Indirect immunofluorescence tests performed on virus-infected MDCK cells showed that this Fab fragment was at least equally efficient as other commercial monoclonal antibody-based systems in detecting influenza A viral infections. The potential advantages of human recombinant Fabs on murine monoclonal antibodies are discussed.

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