Role of pentamer complex-specific and IgG subclass 3 antibodies in HCMV hyperimmunoglobulin and standard intravenous IgG preparations
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HCMV hyperimmunoglobulin-preparations (HIG) contain high concentrations of HCMV-specific IgG. The reduced maternofetal-HCMV-transmission rate of IgG may be due to HCMV-specific neutralizing antibodies against the HCMV pentameric complex (PC). In contrast to HIG, standard intravenous immunoglobulin (IVIG) may have more neutralization (NT) capacity than HIG due to higher IgG subclass 3 levels (Planitzer et al., 2011).
We investigated the HCMV-specific NT-capacity of HIG Cytotect®, using a recombinant pentameric complex (gHgLUL128-131A) for specific antibody-depletion. We used a modified UL130-peptide (TANQNPSPPWSKLTYSKPH) based on original-sequence of Saccoccio et al. (Vaccine 29(15):2705–2711, 2011) (SWSTLTANQNPSPPWSKLTY) as neutralization target. Both UL130-peptides and the PC were bound via sixfold HisTag and anti-HisTag mAbs to magnetic beads to deplete HCMV-specific IgGs from HIG (Cytotect®). Modifying this depletion strategy, we analyzed the role of IgG subclass 3 in both HIG and IVIG.
After CMV IgG-normalization of HIG and IVIG, we found a significant trend towards a decrease (16%) of neutralization-capacity for the UL130 TAN-peptide, but not for the original UL130 SWS-peptide. However, highly significant loss of NT-capacity could be only observed by PC depletion (42%). The IgG subclass 3 depletion revealed no significant reduction of NT-capacity in both HIG and IVIG.
Via specific antibody depletion, we could demonstrate that pentameric complex-specific antibodies are present in HIG and bind to the recombinant PC resulting in a highly significant reduction of NT-capacity compared to the UL130 TAN-and SWS-peptides. We could not confirm the functional role of IgG subclass 3 neutralizing antibodies in IgG-preparations.
KeywordsCytomegalovirus CMV Hyperimmunoglobulin HIG Pentameric complex gHgLUL128-131 Neutralizing antibodies IgG subclass 3
MS received a Grant from Biotest AG, Preclinical Research (Dr. M Germer). Cytotect® was provided from Biotest AG. We thank Wioleta Kapis for her excellent technical assistance and for providing sera from the Tuebingen congenital CMV study.
Compliance with ethical standards
Conflict of interest
All authors declare that they have no conflict of interest.
Maternal sera at birth were derived from the Tuebingen congenital CMV study, approved by our local Ethics Committee at the University Hospital of Tuebingen; EK number 506/2015BO2. Each mother enrolled in the cCMV study was informed about the study design and has given signed permission using her serum for CMV serology.
- 5.Ciferri C, Chandramouli S, Donnarumma D, Nikitin PA, Cianfrocco MA, Gerrein R, Feire AL, Barnett SW1, Lilja AE, Rappuoli R, Norais N, Settembre EC, Carfi A (2015) Structural and biochemical studies of HCMV gH/gL/gO and Pentamer reveal mutually exclusive cell entry complexes. Proc Natl Acad Sci USA 112(6):1767–1772CrossRefGoogle Scholar
- 10.Hahn G, Revello MG, Patrone M, Percivalle E, Campanini G, Sarasini A, Wagner M, Gallina A, Milanesi G, Koszinowski U, Baldanti F, Gerna G (2004) Human cytomegalovirus UL131-128 genes are indispensable for virus growth in endothelial cells and virus transfer to leukocytes. J Virol 78(18):10023–10033CrossRefPubMedCentralGoogle Scholar
- 13.Sinzger C, Eberhardt K, Cavignac Y, Weinstock C, Kessler T et al (2006) Macrophage cultures are susceptible to lytic productive infection by endothelial-cell-propagated human cytomegalovirus strains and present viral IE1 protein to CD4 + T cells despite late downregulation of MHC class II molecules. J Gen Virol 87:1853–1862CrossRefGoogle Scholar
- 16.Ha S, Li F, Troutman MC, Freed DC, Tang A, Loughney JW, Wang D, Wang IM, Vlasak J, Nickle DC, Rustandi RR, Hamm M, DePhillips PA, Zhang N, McLellan JS, Zhu H, Adler SP, McVoy MA, An Z, Fu TM (2017) Neutralization of diverse human cytomegalovirus strains conferred by antibodies targeting viral gH/gL/pUL128-131 pentameric complex. J Virol 91(7):e02033-16. https://doi.org/10.1128/JVI.02033-16 CrossRefPubMedCentralGoogle Scholar
- 17.Zydek M, Petitt M, Fang-Hoover J, Adler B, Kauvar LM, Pereira L, Tabata T (2014) HCMV infection of human trophoblast progenitor cells of the placenta is neutralized by a human monoclonal antibody to glycoprotein B and not by antibodies to the pentamer complex. Viruses 6(3):1346–1364CrossRefPubMedCentralGoogle Scholar
- 20.Visentin S, Manara R, Milanese L, Da Roit A, Forner G, Salviato E, Citton V, Magno FM, Orzan E, Morando C, Cusinato R, Mengoli C, Palu G, Ermani M, Rinaldi R, Cosmi E, Gussetti N (2012) Early primary cytomegalovirus infection in pregnancy: maternal hyperimmunoglobulin therapy improves outcomes among infants at 1 year of age. Clin Infect Dis 55(4):497–503CrossRefGoogle Scholar
- 21.Buxmann H, Stackelberg OM, Schlößer RL, Enders G, Gonser M, Meyer-Wittkopf M, Hamprecht K, Enders M (2012) Use of cytomegalovirus hyperimmunoglobulin for prevention of congenital cytomegalovirus disease: a retrospective analysis. J Perinat Med 40(4):439–446Google Scholar
- 22.Revello MG, Lazzarotto T, Guerra B, Spinillo A, Ferrazzi E, Kustermann A, Guaschino S, Vergani P, Todros T, Frusca T, Arossa A, Furione M, Rognoni V, Rizzo N, Gabrielli L, Klersy C, Gerna G, CHIP Study Group (2014) A randomized trial of hyperimmune globulin to prevent congenital. N Engl J Med 370(14):1316–1326CrossRefGoogle Scholar
- 23.Rawlinson WD, Boppana SB, Fowler KB, Kimberlin DW, Lazzarotto T, Alain S, Daly K, Doutré S, Gibson L, Giles ML, Greenlee J, Hamilton ST, Harrison GJ, Hui L, Jones CA, Palasanthiran P, Schleiss MR, Shand MW, van Zuylen WJ (2017) Congenital cytomegalovirus infection in pregnancy and the neonate: consensus recommendations for prevention, diagnosis, and therapy. Lancet Infect Dis 17(6):e177–e188CrossRefGoogle Scholar
- 24.Kagan KO, Enders M, Schampera MS, Baeumel E, Hoopmann M, Geipel A, Berg C, Goelz R, De Catte L, Wallwiener D, Brucker S, Adler SP, Jahn G, Hamprecht K (2018) Prevention of maternal-fetal transmission of CMV by hyperimmunoglobulin (HIG) administered after a primary maternal CMV infection in early gestation. Ultrasound Obstet Gynecol. https://doi.org/10.1002/uog.19164 Google Scholar
- 25.Schampera MS, Schweinzer K, Abele H, Kagan KO, Klein R, Rettig I, Jahn G, Hamprecht K (2017) Comparison of cytomegalovirus (CMV)-specific neutralization capacity of hyperimmunoglobulin (HIG) versus standard intravenous immunoglobulin (IVIG) preparations: impact of CMV IgG normalization. J Clin Virol 90:40–45CrossRefGoogle Scholar
- 28.Saccoccio FM, Sauer AL, Cui X, Armstrong AE, Habib el-SE, Johnson DC, Ryckman BJ, Klingelhutz AJ, Adler SP, McVoy MA (2011) Peptides from cytomegalovirus UL130 and UL131 proteins induce high titer antibodies that block viral entry into mucosal epithelial cells. Vaccine 29(15):2705–2711CrossRefPubMedCentralGoogle Scholar
- 32.Saha S, Raghava GPS (2004) BcePred: prediction of continuous B-cell epitopes in antigenic sequences using physico-chemical properties, vol 3239. ICARIS, LNCS, Springer, New York, pp 197–204Google Scholar
- 38.Loughney JW, Rustandi RR, Wang D, Troutman MC, Dick LW Jr, Li G, Liu Z, Li F, Freed DC, Price CE, Hoang VM, Culp TD, DePhillips PA, Fu TM, Ha S (2015) Soluble human cytomegalovirus gH/gL/pUL128-131 pentameric complex, but not gH/gL, inhibits viral entry to epithelial cells and presents dominant native neutralizing epitopes. J Biol Chem 290(26):15985–15995CrossRefPubMedCentralGoogle Scholar
- 39.Macagno A, Bernasconi NL, Vanzetta F, Dander E, Sarasini A, Revello MG, Gerna G, Sallusto F, Lanzavecchia A (2010) Isolation of human monoclonal antibodies that potently neutralize human cytomegalovirus infection by targeting different epitopes on the gH/gL/UL128-131A complex. J Virol 84(2):1005–1013CrossRefGoogle Scholar
- 40.Gerna G, Percivalle E, Perez L, Lanzavecchia A, Lilleri D (2016) Monoclonal antibodies to different components of the human cytomegalovirus (HCMV) pentamer gH/gL/pUL128L and trimer gH/gL/gO as well as antibodies elicited during primary HCMV Infection prevent epithelial cell syncytium formation. J Virol 90(14):6216–6223CrossRefPubMedCentralGoogle Scholar
- 41.Ha S, Li F, Troutman MC, Freed DC, Tang A, Loughney JW, Wang D, Wang I-Ming, Vlasak J, Nickle DC, Rustandi RR, Hamm M, DePhillips PA, Zhang N, McLellan JS, Zhu H, Adler SP, McVoy MA, Zhiqiang AN, Tong-Ming FU (2017) Neutralization of diverse human cytomegalovirus strains conferred by antibodies targeting viral gH/gL/pUL128-131 pentameric complex. J Virol 91(7):JVI-02033CrossRefGoogle Scholar
- 43.Hamilton ST, van Zuylen W, Shand A, Scott GM, Naing Z, Hall B, Craig ME, Rawlinson WD (2014) Prevention of congenital cytomegalovirus complications by maternal and neonatal treatments: a systematic review. Rev Med Virol. (6):420–433Google Scholar