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EXPERIMENTAL CLINICAL INVESTIGATION

Serum Interferon (IFN)-Neutralizing Antibodies and Bioactivities of IFNs in Patients with Severe Type II Essential Mixed Cryoglobulinemia

Carolina Scagnolari, Milvia Casato, Francesca Bellomi, Francesca De Pisa, Ombretta Turriziani, Rossella Coviello, Maria Rosaria Pirro, Ferdinando Dianzani, Guido Antonelli
Carolina Scagnolari
1Department of Experimental Medicine, Virology Section
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Milvia Casato
2Department of Clinical Medicine, Policlinico Umberto I
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Francesca Bellomi
1Department of Experimental Medicine, Virology Section
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Francesca De Pisa
3Department of Experimental Medicine, University “La Sapienza”
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Ombretta Turriziani
1Department of Experimental Medicine, Virology Section
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Rossella Coviello
2Department of Clinical Medicine, Policlinico Umberto I
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Maria Rosaria Pirro
2Department of Clinical Medicine, Policlinico Umberto I
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Ferdinando Dianzani
4“Campus Biomedico,” Libera Università, Rome, Italy
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Guido Antonelli
1Department of Experimental Medicine, Virology Section
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  • For correspondence: guido.antonelli@uniroma1.it
DOI: 10.1128/CDLI.10.1.70-77.2003
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  • FIG. 1.
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    FIG. 1.

    Treatment and blood sampling sequences during the study.

  • FIG. 2.
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    FIG. 2.

    QC PCR analysis of MxA mRNA in PBMCs from patient 1. PBMCs were collected before and at 24 and 48 h after a single injection of rIFN-a2a, (C)IFN, and LeIFN. Serial logarithmic dilutions of extracted RNA, normalized by using b-actin mRNA as a control, were mixed with 2 3 106 copies of the internal competitor and subjected to RT-PCR (for details, see Materials and Methods). The arrows indicate the equivalence points.

  • FIG. 3.
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    FIG. 3.

    QC PCR analysis of MxA mRNA in PBMCs from patient 2. PBMCs were collected before and at 24 and 48 h after a single injection of rIFN-α2a, (C)IFN, and LeIFN. Serial logarithmic dilutions of extracted RNA, normalized by using β-actin mRNA as a control, were mixed with 2 × 106 copies of the internal competitor and subjected to RT-PCR (for details, see Materials and Methods). The arrows indicate the equivalence points.

  • FIG. 4.
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    FIG. 4.

    Neopterin protein levels in the sera of patients 1 (A) and 2 (B). Serum neopterin concentrations were measured by enzyme-linked immunosorbent assay before and at 24 and 48 h after a single injection of rIFN-α2a, (C)IFN, and LeIFN. Bars represent means ± standard deviations for three separate determinations.

  • FIG. 5.
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    FIG. 5.

    Increases in serum neopterin levels at 24 and 48 h compared with that at the baseline in NAB-negative patients with EMC after a single injection of recombinant IFN-α2a. Each histogram represents the mean ± standard deviations for five patients. ∗, P < 0.05 compared with the level at time zero.

Tables

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  • TABLE 1.

    Neutralizing activities of sera from two patients with severe type II EMC resistant to rIFN-α2aa

    Patient no.Neutralization titer against:
    rIFN-α2a(C)IFNLeIFN
    1b950 ± 19063 ± 10NDc
    1d486 ± 8030 ± 5ND
    1e1379 ± 2987 ± 4<10
    2b47 ± 11<10ND
    2d1,300 ± 26087 ± 20ND
    2e1,600 ± 39987 ± 2<10
    • ↵ a Sera for detection of neutralizing activity were collected before the injection of the different IFN-α preparations (see text for details). The data are expressed as the means ± standard deviations for three separate determinations.

    • ↵ b Serum was analyzed before the injection of rIFN-α2a.

    • ↵ c ND, not done.

    • ↵ d Serum was analyzed before the first injection of (C)IFN.

    • ↵ e Serum was analyzed before the first injection of LeIFN.

  • TABLE 2.

    Specificities of NABs to IFN-α in sera from two patients with severe type II EMC resistant to rIFN-α2aa

    IFN-α subtypeNeutralizing activity (t1/10) against subtypes of natural IFN-α
    Patient 1Patient 2
    124 ± 8<10
    2a1,379 ± 291,600 ± 399
    5112 ± 64108 ± 3
    61,124 ± 11189 ± 8
    7a14 ± 533 ± 11
    7c83 ± 53136 ± 76
    8b<10<10
    10a14 ± 349 ± 20
    14a<1084 ± 7
    17a37 ± 243 ± 4
    21a22 ± 13<10
    • ↵ a Sera for detection of the neutralizing activity against the different IFN-α preparations were collected before the injection of LeIFN (see text for details). The data are expressed as the means ± standard deviations for three separate experiments.

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Serum Interferon (IFN)-Neutralizing Antibodies and Bioactivities of IFNs in Patients with Severe Type II Essential Mixed Cryoglobulinemia
Carolina Scagnolari, Milvia Casato, Francesca Bellomi, Francesca De Pisa, Ombretta Turriziani, Rossella Coviello, Maria Rosaria Pirro, Ferdinando Dianzani, Guido Antonelli
Clinical and Diagnostic Laboratory Immunology Jan 2003, 10 (1) 70-77; DOI: 10.1128/CDLI.10.1.70-77.2003

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Serum Interferon (IFN)-Neutralizing Antibodies and Bioactivities of IFNs in Patients with Severe Type II Essential Mixed Cryoglobulinemia
Carolina Scagnolari, Milvia Casato, Francesca Bellomi, Francesca De Pisa, Ombretta Turriziani, Rossella Coviello, Maria Rosaria Pirro, Ferdinando Dianzani, Guido Antonelli
Clinical and Diagnostic Laboratory Immunology Jan 2003, 10 (1) 70-77; DOI: 10.1128/CDLI.10.1.70-77.2003
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