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Immune Repertoire Diversity Correlated with Mortality in Avian Influenza A (H7N9) Virus Infected Patients

View Article: PubMed Central - PubMed

ABSTRACT

Specific changes in immune repertoires at genetic level responding to the lethal H7N9 virus are still poorly understood. We performed deep sequencing on the T and B cells from patients recently infected with H7N9 to explore the correlation between clinical outcomes and immune repertoire alterations. T and B cell repertoires display highly dynamic yet distinct clonotype alterations. During infection, T cell beta chain repertoire continues to contract while the diversity of immunoglobulin heavy chain repertoire recovers. Patient recovery is correlated to the diversity of T cell and B cell repertoires in different ways – higher B cell diversity and lower T cell diversity are found in survivors. The sequences clonally related to known antibodies with binding affinity to H7 hemagglutinin could be identified from survivors. These findings suggest that utilizing deep sequencing may improve prognostication during influenza infection and could help in development of antibody discovery methodologies for the treatment of virus infection.

No MeSH data available.


Related in: MedlinePlus

Identification of broadly neutralizing antibody (bnAb) and H7N9 specific antibody in Ig repertoires of H7N9 infected patients.(a) Alignment of previously reported bnAb sequence (FI390) and the antibody sequence found in H7N9 patient. FI390 was as a bnAb against influenza virus reported by Pappas L. N2_3050 was the IGH sequence found in two recovered H7N9 patients with identical CDR3 amino acid sequence as FI390. Alignment was performed using Clustal W2. (b) Binding kinetics of scFvs N2_3050, o1b1 and l1b1 to H7N9 HA antigen using Surface Plasmon Resonance assays. Different concentrations (10 uM, 2 uM, 400 nM, 200 nM, 100 nM and 50 nM for N2_3050 and c1b1; 1 uM, 500 nM, 250 nM, 125 nM, 62.5 nM and 31.5 nM for l1b1, respectively) is used in the test. Calculated KD values are shown in the table presenting binding affinity of each scFvs.
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f6: Identification of broadly neutralizing antibody (bnAb) and H7N9 specific antibody in Ig repertoires of H7N9 infected patients.(a) Alignment of previously reported bnAb sequence (FI390) and the antibody sequence found in H7N9 patient. FI390 was as a bnAb against influenza virus reported by Pappas L. N2_3050 was the IGH sequence found in two recovered H7N9 patients with identical CDR3 amino acid sequence as FI390. Alignment was performed using Clustal W2. (b) Binding kinetics of scFvs N2_3050, o1b1 and l1b1 to H7N9 HA antigen using Surface Plasmon Resonance assays. Different concentrations (10 uM, 2 uM, 400 nM, 200 nM, 100 nM and 50 nM for N2_3050 and c1b1; 1 uM, 500 nM, 250 nM, 125 nM, 62.5 nM and 31.5 nM for l1b1, respectively) is used in the test. Calculated KD values are shown in the table presenting binding affinity of each scFvs.

Mentions: To examine whether the highly diverse IGH repertoires in some H7N9 patients correlate with the presence of neutralizing antibodies, we firstly compared our deep sequencing data to the reported influenza bnAbs sequences. Surprisingly, we found a panel of clones that have almost identical IGH CDR3 with a previously reported pan-influenza A neutralizing antibody, FI39021. As shown in Fig. 6a, one of these clones, N2_3050, shared ~92% identity with FI390, with a difference of only seven residues in all three CDR regions. This convergence is remarkable considering that IGH repertoires are very diverse among different people, and not a single shared IGH CDR3 sequence can be found in the H7N9 patients investigated here. All these clones were from the samples of two survivors, collected during the convalescent phase (P2, N2). These results suggest that bnAbs may have been elicited in some patients.


Immune Repertoire Diversity Correlated with Mortality in Avian Influenza A (H7N9) Virus Infected Patients
Identification of broadly neutralizing antibody (bnAb) and H7N9 specific antibody in Ig repertoires of H7N9 infected patients.(a) Alignment of previously reported bnAb sequence (FI390) and the antibody sequence found in H7N9 patient. FI390 was as a bnAb against influenza virus reported by Pappas L. N2_3050 was the IGH sequence found in two recovered H7N9 patients with identical CDR3 amino acid sequence as FI390. Alignment was performed using Clustal W2. (b) Binding kinetics of scFvs N2_3050, o1b1 and l1b1 to H7N9 HA antigen using Surface Plasmon Resonance assays. Different concentrations (10 uM, 2 uM, 400 nM, 200 nM, 100 nM and 50 nM for N2_3050 and c1b1; 1 uM, 500 nM, 250 nM, 125 nM, 62.5 nM and 31.5 nM for l1b1, respectively) is used in the test. Calculated KD values are shown in the table presenting binding affinity of each scFvs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC5037391&req=5

f6: Identification of broadly neutralizing antibody (bnAb) and H7N9 specific antibody in Ig repertoires of H7N9 infected patients.(a) Alignment of previously reported bnAb sequence (FI390) and the antibody sequence found in H7N9 patient. FI390 was as a bnAb against influenza virus reported by Pappas L. N2_3050 was the IGH sequence found in two recovered H7N9 patients with identical CDR3 amino acid sequence as FI390. Alignment was performed using Clustal W2. (b) Binding kinetics of scFvs N2_3050, o1b1 and l1b1 to H7N9 HA antigen using Surface Plasmon Resonance assays. Different concentrations (10 uM, 2 uM, 400 nM, 200 nM, 100 nM and 50 nM for N2_3050 and c1b1; 1 uM, 500 nM, 250 nM, 125 nM, 62.5 nM and 31.5 nM for l1b1, respectively) is used in the test. Calculated KD values are shown in the table presenting binding affinity of each scFvs.
Mentions: To examine whether the highly diverse IGH repertoires in some H7N9 patients correlate with the presence of neutralizing antibodies, we firstly compared our deep sequencing data to the reported influenza bnAbs sequences. Surprisingly, we found a panel of clones that have almost identical IGH CDR3 with a previously reported pan-influenza A neutralizing antibody, FI39021. As shown in Fig. 6a, one of these clones, N2_3050, shared ~92% identity with FI390, with a difference of only seven residues in all three CDR regions. This convergence is remarkable considering that IGH repertoires are very diverse among different people, and not a single shared IGH CDR3 sequence can be found in the H7N9 patients investigated here. All these clones were from the samples of two survivors, collected during the convalescent phase (P2, N2). These results suggest that bnAbs may have been elicited in some patients.

View Article: PubMed Central - PubMed

ABSTRACT

Specific changes in immune repertoires at genetic level responding to the lethal H7N9 virus are still poorly understood. We performed deep sequencing on the T and B cells from patients recently infected with H7N9 to explore the correlation between clinical outcomes and immune repertoire alterations. T and B cell repertoires display highly dynamic yet distinct clonotype alterations. During infection, T cell beta chain repertoire continues to contract while the diversity of immunoglobulin heavy chain repertoire recovers. Patient recovery is correlated to the diversity of T cell and B cell repertoires in different ways – higher B cell diversity and lower T cell diversity are found in survivors. The sequences clonally related to known antibodies with binding affinity to H7 hemagglutinin could be identified from survivors. These findings suggest that utilizing deep sequencing may improve prognostication during influenza infection and could help in development of antibody discovery methodologies for the treatment of virus infection.

No MeSH data available.


Related in: MedlinePlus