Human monoclonal antibodies derived from a patient infected with 2009 pandemic influenza A virus broadly cross-neutralize group 1 influenza viruses

Pan, Yang; Sasaki, Tadahiro; Kubota‐Koketsu, Ritsuko; Inoue, Yuji; Yasugi, Mayo; Yamashita, Akifumi; Ramadhany, Ririn; Arai, Yasuha; Du, Anariwa; Boonsathorn, Naphatsawan; Ibrahim, Madiha S.; Daidoji, Tomo; Nakaya, Takaaki; Ono, Kenichiro; Okuno, Yoshinobu; Ikuta, Kazuyoshi; Watanabe, Yohei

doi:10.1016/j.bbrc.2014.05.060

Cited by 11 publications

(6 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, we have reported a cell-to-cell fusion method using a new fusion partner cell line, SPYMEG, and donor-derived peripheral blood mononuclear cells (PBMCs) [9,[15][16][17][18]. In this study, we report the characterization of an anti-influenza HuMAb generated from the PBMCs of a volunteer vaccinated with seasonal influenza vaccine.…”

Section: Introductionmentioning

confidence: 65%

“…Total RNA was extracted from the hybridoma cells using an RNeasy Mini Kit (Qiagen) and subjected to RT-PCR as described previously [9]. The specific PCR product was then sequenced and the sequences obtained were used to search the NCBI database using the IgBLAST website (http://www.ncbi.nlm.nih.gov/igblast/).…”

Section: Sequencing Of the Humab Variable Regionsmentioning

confidence: 99%

“…Furthermore, the HA head region rapidly and continuously accumulates amino acid substitutions to escape from the antibody response. Recently, several anti-stem region antibodies with a broad spectrum against influenza viruses have been reported [7][8][9][10]. However, information about conserved HA epitopes is still limited.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A human monoclonal antibody derived from a vaccinated volunteer recognizes heterosubtypically a novel epitope on the hemagglutinin globular head of H1 and H9 influenza A viruses

Boonsathorn

Panthong

Koksunan

et al. 2014

Biochemical and Biophysical Research Communications

Self Cite

View full text Add to dashboard Cite

Most neutralizing antibodies elicited during influenza virus infection or by vaccination have a narrow spectrum because they usually target variable epitopes in the globular head region of hemagglutinin (HA). In this study, we describe a human monoclonal antibody (HuMAb), 5D7, that was prepared from the peripheral blood lymphocytes of a vaccinated volunteer using the fusion method. The HuMAb heterosubtypically neutralizes group 1 influenza A viruses, including seasonal H1N1, 2009 pandemic H1N1 (H1N1pdm) and avian H9N2, with a strong hemagglutinin inhibition activity. Selection of an escape mutant showed that the HuMAb targets a novel conformational epitope that is located in the HA head region but is distinct from the receptor binding site. Furthermore, Phe114Ile substitution in the epitope made the HA unrecognizable by the HuMAb. Amino acid residues in the predicted epitope region are also highly conserved in the HAs of H1N1 and H9N2. The HuMAb reported here may be a potential candidate for the development of therapeutic/prophylactic antibodies against H1 and H9 influenza viruses.

show abstract

Section: Introductionmentioning

confidence: 65%

Section: Sequencing Of the Humab Variable Regionsmentioning

confidence: 99%

See 1 more Smart Citation

A human monoclonal antibody derived from a vaccinated volunteer recognizes heterosubtypically a novel epitope on the hemagglutinin globular head of H1 and H9 influenza A viruses

Boonsathorn

Panthong

Koksunan

et al. 2014

Biochemical and Biophysical Research Communications

Self Cite

View full text Add to dashboard Cite

show abstract

“…A very unique heterohybridoma fusion partner denoted SPYMEG was developed by fusion of Sp2 murine myeloma cells and MEG-01 human megakaryoblastic leukemia cells (74). Recently, SPYMEG was employed for the production of anti-influenza mAbs, generated by fusing PBMCs from influenza-vaccinated and naturally infected volunteers (75,76). It was also used successfully to make human hybridomas secreting anti-HIV neutralizing mAbs by fusion with PBMCs obtained from HIV-1-infected individuals (77).…”

Section: Fusion Partnersmentioning

confidence: 99%

Use of Human Hybridoma Technology To Isolate Human Monoclonal Antibodies

Smith

Crowe

2015

Microbiol Spectr

View full text Add to dashboard Cite

The human hybridoma technique offers an important approach for isolation of human monoclonal antibodies. A diversity of approaches can be used with varying success. Recent technical advances in expanding the starting number of human antigen-specific B cells, improving fusion efficiency, and isolating new myeloma partners and new cell cloning methods have enabled the development of protocols that make the isolation of human monoclonal antibodies from blood samples feasible. Undoubtedly, additional innovations that could improve efficiency are possible. HISTORY OF HYBRIDOMASMonoclonal antibodies (mAbs) have revolutionized the conduct of science since their first description in 1975 (1). The use of these specific reagents also has made possible improved clinical diagnostics in the medical arena, and many antibodies have found their way to clinical use as prophylactic or therapeutic agents. Nevertheless, the potential of mAbs derived specifically from technology based on human hybridomas remains largely unfulfilled. The principal reason for the lack of a large number of hybridoma-derived mAb therapeutics has simply been the technical difficulty in generating stable hybridomas that secrete human mAbs of high affinity and functional activity. This chapter reviews recent efforts to develop and employ novel methods for the efficient generation of human hybridomas secreting human mAbs for clinical use.The principal advantage of the use of human hybridoma technology for mAb generation is that this approach preserves the authentic sequence and pairing of antibody DNA from a natural B cell for the expression of a naturally occurring full-length human mAb. There are significant theoretical advantages for expressing cDNAs encoding authentic heavy and light chains with chains that are paired using the coding sequence as it was generated naturally through B cell selection, class switch, and affinity maturation. No genetic modification of these sequences is required. Since antibody expression is typically very stable in hybridomas, sequence amplification of antibody variable genes is achieved easily if recombinant production or manipulation is desired. The resulting recombinant mAb retains most features of naturally occurring human antibodies, as the clone retains the native amino acid sequence and heavy/light chain pairing. Since the native constant region of the antibody in the original human B cell is retained in the mAb expressed by the resulting hybridoma, the functional properties of the particular Fc region can be studied for Fc-mediated activities, such as antibodydependent cellular cytotoxicity.

show abstract

“…SPYMEG was optimized from a human megakaryoblastic leukemia cell line (MEG-01), obtained from a patient with blast crisis of Philadelphia chromosome-positive chronic myelogenous leukemia, via fusion with SP2 murine myeloma cells [ 10 , 19 ]. SPYMEG has been successfully used as a fusion partner to obtain mAbs that recognize the HA of influenza A [ 10 , 14 , 20 ] and influenza B viruses [ 28 ], and the dengue virus envelope protein [ 23 , 24 ]. However, as with most partner cell lines, little information regarding the efficiency of SPYMEG has been reported.…”

mentioning

confidence: 99%

Evaluation of the fusion partner cell line SPYMEG for obtaining human monoclonal antibodies against influenza B virus

Soni

Yasuhara

Takenaga

et al. 2018

The Journal of Veterinary Medical Science

Self Cite

View full text Add to dashboard Cite

Influenza B virus has been known to infect humans and other animals, including seals. Vaccination efficacy varies across seasons. Human monoclonal antibodies (mAbs) can be useful for developing novel vaccines, guided by epitope analysis, and can be used therapeutically. Hybridoma technology has been used to make mAbs. Here we evaluated SPYMEG as a fusion partner cell line for human mAb generation specific to influenza B hemagglutinin (HA). SPYMEG is a human/murine myeloma partner cell line that has previously been used to generate human mAbs that recognize the HA of influenza A and B viruses. Peripheral blood mononuclear cells were obtained from 16 volunteers, previously vaccinated with the 2014–2015 trivalent seasonal influenza vaccine, and were fused with SPYMEG to yield hybridomas. The resulting hybridomas were screened for antigen-specific antibody secretion and cloned by limiting dilution. We obtained 32 stable clones secreting anti-influenza B HA human IgG, although most of these clones were obtained from one volunteer (SeaV-29) who had a robust immune response. We conclude that SPYMEG is a good fusion partner cell line, although cloning by limiting dilution may lead to significant loss of hybridomas.

show abstract

Human monoclonal antibodies derived from a patient infected with 2009 pandemic influenza A virus broadly cross-neutralize group 1 influenza viruses

Cited by 11 publications

References 27 publications

A human monoclonal antibody derived from a vaccinated volunteer recognizes heterosubtypically a novel epitope on the hemagglutinin globular head of H1 and H9 influenza A viruses

A human monoclonal antibody derived from a vaccinated volunteer recognizes heterosubtypically a novel epitope on the hemagglutinin globular head of H1 and H9 influenza A viruses

Use of Human Hybridoma Technology To Isolate Human Monoclonal Antibodies

Evaluation of the fusion partner cell line SPYMEG for obtaining human monoclonal antibodies against influenza B virus

Contact Info

Product

Resources

About