Region Specific and Worldwide Distribution of Collagen-Binding M Proteins with PARF Motifs among Human Pathogenic Streptococcal Isolates

Some of the variety of Streptococcus pyogenes and Streptococcus dysgalactiae ssp. equisimilis (SDSE) M proteins act as collagen-binding adhesins that facilitate acute infection. Moreover, their potential to trigger collagen autoimmunity has been implicated in the pathogenesis of acute rheumatic fever and attributed to a collagen-binding motif called PARF (peptide associated with rheumatic fever). For the first time we determine the rate of clinical isolates with collagen-binding M proteins that use a PARF motif (A/T/E)XYLXX(L/F)N in a defined geographic region, Vellore in South India. In this region both, incidence of streptococcal infections and prevalence of acute rheumatic fever are high. M proteins with PARF motif conferred collagen-binding activity to 3.9% of 153 S. pyogenes and 10.6% of 255 SDSE clinical isolates from Vellore. The PARF motif occurred in three S. pyogenes and 22 SDSE M protein types. In one of the S. pyogenes and five of the SDSE M proteins that contained the motif, collagen-binding was impaired, due to influences of other parts of the M protein molecule. The accumulated data on the collagen binding activity of certain M protein types allowed a reanalysis of published worldwide emm-typing data with the aim to estimate the rates of isolates that bind collagen via PARF. The results indicate that M proteins, which bind collagen via a PARF motif, are epidemiologically relevant in human infections, not only in Vellore. It is imperative to include the most relevant collagen-binding M types in vaccines. But when designing M protein based vaccines it should be considered that collagen binding motifs within the vaccine antigen remain potential risk factors

Contribution of Streptococcus anginosus to Infections Caused by Groups C and G Streptococci, Southern India

This neglected pathogen causes a large portion of these infections

Human C1q Regulates Influenza A Virus Infection and Inflammatory Response via Its Globular Domain

The Influenza A virus (IAV) is a severe respiratory pathogen. C1q is the first subcomponent of the complement system’s classical pathway. C1q is composed of 18 polypeptide chains. Each of these chains contains a collagen-like region located at the N terminus, and a C-terminal globular head region organized as a heterotrimeric structure (ghA, ghB and ghC). This study was aimed at investigating the complement activation-independent modulation by C1q and its individual recombinant globular heads against IAV infection. The interaction of C1q and its recombinant globular heads with IAV and its purified glycoproteins was examined using direct ELISA and far-Western blotting analysis. The effect of the complement proteins on IAV replication kinetics and immune modulation was assessed by qPCR. The IAV entry inhibitory properties of C1q and its recombinant globular heads were confirmed using cell binding and luciferase reporter assays. C1q bound IAV virions via HA, NA and M1 IAV proteins, and suppressed replication in H1N1, while promoting replication in H3N2-infected A549 cells. C1q treatment further triggered an anti-inflammatory response in H1N1 and pro-inflammatory response in H3N2-infected cells as evident from differential expression of TNF-α, NF-κB, IFN-α, IFN-β, IL-6, IL-12 and RANTES. Furthermore, C1q treatment was found to reduce luciferase reporter activity of MDCK cells transfected with H1N1 pseudotyped lentiviral particles, indicative of an entry inhibitory role of C1q against infectivity of IAV. These data appear to demonstrate the complement-independent subtype specific modulation of IAV infection by locally produced C1q

<i>Emm-t</i>yping of clinical <i>S. pyogenes</i> isolates from Vellore.

a<p>emm3.22 (3 isolates); emm3.23 (3 isolates).</p

Prediction of coiled-coil structures in PARF-positive M proteins.

<p>Coiled-coil structure prediction for the N-terminal sequences of the PARF-positive M proteins stG120.0 (<i>red dotted line</i>), stG120.1 (<i>light blue solid line</i>) and stGM220 (<i>black dotted line</i>) is given as P-scores vs. the amino acid position relative from the PARF motif with the first amino acid of the motif being position 1. Isolates with M protein stG120.1 bind collagen IV, while strains that carry one of the other two M proteins do not. The position of the PARF motif is highlighted in <i>light blue</i>. A <i>black dashed line</i> indicates the threshold value for coiled-coil prediction. Values below 0.025 indicate a coiled-coil structure.</p

Prediction of N-terminal coiled-coil structures in PARF-positive M proteins.

<p>(<i>A</i>) Coiled-coil structure prediction for the N-terminal sequences of the PARF-positive M proteins is given as P-scores vs. the amino acid position relative from the PARF motif with the first amino acid of the motif being position 1. The figure shows the superposition of all curves without indicating protein designations, however, collagen-binding M proteins are shown in <i>blue</i> and non-binding M proteins are shown in <i>red</i>. Single curves for all M proteins are provided as a supplementary figure (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030122#pone.0030122.s001" target="_blank">Figure S1</a>). The position of the PARF motif is highlighted in <i>light blue</i>. A <i>black dashed line</i> indicates the threshold value for coiled-coil prediction. Values below 0.025 indicate a coiled-coil structure. The prediction separates the M proteins into two classes; one class with (<i>B</i>) and one without (<i>C</i>) an N-terminal coiled-coil domain. They are shown in schematic representations that are not drawn to scale. <i>B</i> and <i>C</i> are not based on structural data other than coiled-coil prediction. The PARF motif is highlighted in <i>red</i>. The C-terminal end is labelled (-COOH).</p

M proteins with predicted PARF motifs that did not bind collagen.

<p>Protein sequences of four types or groups of M proteins that harbored a prototypical PARF motif but had members that did not bind collagen (<i>A–D</i>). M protein designations are given on the left together with the results of collagen IV binding experiments that are given in brackets with ‘+’ for binding and ‘−’ for non-binding proteins. Numbers that flank the sequence indicate its position within the mature M protein without signal peptide. In <i>C</i> and <i>D</i> identical amino acids (*), conserved substitutions (:), and semi-conserved substitutions (.) in two or three compared sequences are marked below. Amino acids characteristic for PARF are printed in <i>bold</i>. In <i>D</i> residues that distinguish the collagen-IV binding stG120.1 from the non-binding stG120.0 and stGM220 are highlighted in <i>grey</i>.</p

PARF-like motifs found in M proteins of <i>S. pyogenes</i> and <i>S. dysgalactiae</i> subsp. <i>equisimilis</i> isolated in Vellore.

a<p>Amino acids conserved or typical for PARF are shown in bold.</p>b<p>Proteins M110 and M92 harbored two PARF-like motifs.</p