Research ArticleINFECTIOUS DISEASE

Recovery from the Middle East respiratory syndrome is associated with antibody and T cell responses

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Science Immunology  04 Aug 2017:
Vol. 2, Issue 14, eaan5393
DOI: 10.1126/sciimmunol.aan5393
  • Fig. 1 Convalescent sera transfer protects mice from MERS-CoV infection.

    (A) Mice received 75 μl of patient serum intravenously 12 hours before MERS-CoV infection. One hour before infection, mice sera were collected and PRNT50 assays were performed as described in Materials and Methods. HD, healthy donor; LOD, limit of detection. (B) Relationship between PRNT50 in human sera and in mouse recipients of transferred sera. (C) To obtain virus titers, we homogenized lungs at day 3 after infection and titered on Vero 81 cells. Titers are expressed as PFU/g of tissue. n = 3 mice per group per time point. Right: Relationship between PRNT50 in mouse sera and viral titers in mouse lungs. AT, adoptive transfer.

  • Fig. 2 Virus-specific T cell responses are detected in all MERS survivors.

    PBMCs from healthy donors and MERS patients were stimulated with MERS-CoV structural protein–specific peptide pools for 12 hours in the presence of brefeldin A. Frequencies of MERS-CoV–specific CD4+ (A and B) and CD8+ (C and D) T cells (determined by IFN-γ intracellular staining) are shown. (E) Summary of total T cell responses against all four peptide pools is shown. (F) Relationship between T cell and neutralizing antibody responses is shown.

  • Fig. 3 Human PBMC-derived MERS-CoV–specific T cells are multifunctional.

    (A and C) PBMCs were stimulated with MERS-CoV structural protein–specific peptide pools. Frequency and percentage of cells expressing IFN-γ and TNF are shown. (B and D) PBMCs were stimulated with the N (B) or ME (D) peptide pools. CD4+ (B) or CD8+ (D) T cells were then analyzed for the indicated phenotypic markers.

  • Fig. 4 Identification of MERS-CoV–specific T cell epitopes in mice and patients.

    (A) Single-cell suspensions were prepared from the lungs of MERS-CoV–infected DR2 and DR3 transgenic mice and stimulated with peptides for 5 to 6 hours in the presence of brefeldin A. (B to D) DR2- or DR3-restricted patient PBMCs were stimulated with peptide pools or individual peptides for 12 hours in the presence of brefeldin A. (E) Patient PBMCs were stimulated with the ME peptide pool or individual peptides for 12 hours in the presence of brefeldin A. Frequencies of MERS-CoV–specific T cells (determined by IFN-γ intracellular staining) are shown.

  • Fig. 5 Relationship between MERS-CoV–specific T cell and neutralizing antibody responses and disease variables and severity.

    (A) Relationship between T cell and PRNT50 responses and time after infection when samples were obtained. Mon, months; n.s., not significant. (B and C) Relationship between T cell (B) and PRNT50 (C) responses and comorbidity (comorbidity versus none), ventilator status, sex, and age. (D and E) Relationship between T cell and PRNT50 responses and the duration of virus shedding (D) and length of ICU stay (E).

  • Table 1 Serological testing.

    Red, severe pneumonia; blue, pneumonia; green, asymptomatic; black, PMBCs not available; N.D., not done.

    PT IDELISA resultELISAIFAIFA titerMicroneutralization titerPRNT50
    PT01Positive12.3Positive10063.51057
    PT03Negative0.23Negative<1:10≤10≤20
    PT05Positive6.1Positive100226.31432
    PT08Positive4.1Positive100100.8592
    PT09Positive4.93Positive100226.31170
    PT10Positive5.1Positive100201.6912
    PT11Positive2.3Positive1:1025.2148
    PT18Positive2.2Positive10050.4370
    PT19Positive2.98Positive10040278
    PT02Positive2.02Positive1:1080930
    PT04Borderline0.87Positive1:10≤1031
    PT06Positive1.34Positive10015.943.5
    PT07Borderline0.97Negative<1:10≤10128
    PT20Positive4.4Positive1:10 weak40293
    PT21Positive1.17Borderline1:10 weak≤1061
    PT12Negative0.56Negative<1:10≤10≤20
    PT13Negative0.36Negative<1:10≤10≤20
    PT14Negative0.38Negative<1:10≤10≤20
    PT15N.D.N.D.N.D.N.D.28.3200
    PT16Positive3.4Positive100100.8301
    PT17Positive1.85Positive10025.2247

Supplementary Materials

  • Supplementary Materials

    Supplementary Material for:

    Recovery from the Middle East respiratory syndrome is associated with antibody and T cell responses

    Jingxian Zhao, Abeer N. Alshukairi,* Salim A. Baharoon, Waleed A. Ahmed, Ahmad A. Bokhari, Atef M. Nehdi, Laila A. Layqah, Mohammed G. Alghamdi, Manal M. Al Gethamy, Ashraf M. Dada, Imran Khalid, Mohamad Boujelal, Sameera M. Al Johani, Leatrice Vogel, Kanta Subbarao, Ashutosh Mangalam, Chaorong Wu, Patrick Ten Eyck, Stanley Perlman,* Jincun Zhao*

    *Corresponding authors. Email: stanley-perlman{at}uiowa.edu (S.P.); zhaojincun{at}gird.cn (J.Z.); aalshukairi{at}kfshrc.edu.sa (A.N.A.)

    Published 4 August 2017, Sci. Immunol. 2, eaan5393 (2017)
    DOI: 10.1126/sciimmunol.aan5393

    This PDF file includes:

    • Fig. S1. Gating strategy for determining cellular composition of PBMCs.
    • Table S1. Clinical information including laboratory values.
    • Table S2. PBMC composition.
    • Table S3. Peptide list.
    • Table S4. HLA typing.

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    Other Supplementary Material for this manuscript includes the following:

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