Intranasal Administration of Antibody-Bound Respiratory Syncytial Virus Particles Efficiently Primes Virus-Specific Immune Responses in Mice

Kruijsen, Debby; Einarsdóttir, Hildur; Schijf, Marcel A.; Coenjaerts, Frank E. J.; Schoot, Ellen C. van der; Vidarsson, Gestur; Bleek, Grada M. van

doi:10.1128/jvi.00493-13

Cited by 31 publications

(39 citation statements)

References 47 publications

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“…A single-gene vector anti-TNP IgG1 heavy- and kappa-light chain encoding sequences were cloned as described previously by Kruijssen et al 51. into a pEE14.4 (Lonza) expression vector.…”

Section: Methodsmentioning

confidence: 99%

Multi-level glyco-engineering techniques to generate IgG with defined Fc-glycans

Dekkers

Plomp

Koeleman

et al. 2016

Sci Rep

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106

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Immunoglobulin G (IgG) mediates its immune functions through complement and cellular IgG-Fc receptors (FcγR). IgG contains an evolutionary conserved N-linked glycan at position Asn297 in the Fc-domain. This glycan consists of variable levels of fucose, galactose, sialic acid, and bisecting N-acetylglucosamine (bisection). Of these variations, the lack of fucose strongly enhances binding to the human FcγRIII, a finding which is currently used to improve the efficacy of therapeutic monoclonal antibodies. The influence of the other glycan traits is largely unknown, mostly due to lack of glyco-engineering tools. We describe general methods to produce recombinant proteins of any desired glycoform in eukaryotic cells. Decoy substrates were used to decrease the level of fucosylation or galactosylation, glycosyltransferases were transiently overexpressed to enhance bisection, galactosylation and sialylation and in vitro sialylation was applied for enhanced sialylation. Combination of these techniques enable to systematically explore the biological effect of these glycosylation traits for IgG and other glycoproteins.

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“…A single-gene vector anti-TNP IgG1 heavy- and kappa-light chain encoding sequences were cloned as described previously by Kruijssen et al 51. into a pEE14.4 (Lonza) expression vector.…”

Section: Methodsmentioning

confidence: 99%

Multi-level glyco-engineering techniques to generate IgG with defined Fc-glycans

Dekkers

Plomp

Koeleman

et al. 2016

Sci Rep

Self Cite

106

119

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“…Subsequently, the codon-optimized V gene for heavy chain or V gene and k constant region gene for light chain, including 5 0 -HindIII and 3 0 -NheI or 5 0 -HindIII and 3 0 -EcoRI restriction sites, respectively Kozak sequence and HAVT20-leader sequence (Vidarsson et al, 2001), were designed and ordered from Geneart (Life Technologies, Carlsbad, CA, USA). The HindIII-NheI fragment for the codon-optimized heavy chain was ligated into c constant region flanking 3 0 -EcoRI restriction site, as described by (Kruijsen et al, 2013). The HindIII-EcoRI fragment for the codon-optimized light chain was ligated into pEE14.4 (Lonza, Williamsport, PA, USA), and the full HindIII-EcoRI fragment for the heavy chain was ligated into pEE6.4 (Lonza).…”

Section: Recombinant Anti-hpa-1amentioning

confidence: 99%

Glycosylation pattern of anti‐platelet IgG is stable during pregnancy and predicts clinical outcome in alloimmune thrombocytopenia

et al. 2016

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Fetal or neonatal alloimmune thrombocytopenia (FNAIT) is a potentially life-threatening disease where fetal platelets are destroyed by maternal anti-platelet IgG alloantibodies. The clinical outcome varies from asymptomatic, to petechiae or intracranial haemorrhage, but no marker has shown reliable correlation with severity, making screening for FNAIT impractical and highly inefficient. We recently found IgG Fc-glycosylation towards platelet and red blood cell antigens to be skewed towards decreased fucosylation, increased galactosylation and sialylation. The lowered core-fucosylation increases the affinity of the pathogenic antibodies to FcγRIIIa and FcγRIIIb, and hence platelet destruction. Here we analysed the N-linked glycans of human platelet antigen (HPA)-1a specific IgG1 with mass spectrometry in large series of FNAIT cases (n = 166) including longitudinal samples (n = 26). Besides a significant decrease in Fc-fucosylation after the first pregnancy (P = 0·0124), Fc-glycosylation levels remained stable during and after pregnancy and in subsequent pregnancies. Multiple logistic regression analysis identified anti-HPA-1a -fucosylation (P = 0·006) combined with galactosylation (P = 0·021) and antibody level (P = 0·038) correlated with bleeding severity, making these parameters a feasible marker in screening for severe cases of FNAIT.

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“…The variable regions of the heavy and light chains (VH, VL) of the mouse IgG1 anti-2,4,6-trinitrophenol (TNP) hapten antibodies were cloned onto human IgG1 or k backbone, respectively, as described previously, 35 and produced in the HEK-293F FreeStyle cell line expression system (Life Technologies, Paisley, United Kingdom), but now in the presence or absence of 2-deoxy-2-fluoro-L-fucose (2F; Carbosynth, Compton, Berkshire, United Kingdom) to control the level of fucosylation. 36 Antibodies were purified on a protein A (WT IgG1) HiTrap HP column (GE Healthcare Life Sciences, Little Chalfont, United Kingdom) using the Acta Prime Plus system (GE Healthcare Life Sciences) and dialyzed against phosphate-buffered saline (PBS) overnight.…”

Section: Production Of Recombinant Anti-tnp Igg1 Antibodies With Low mentioning

confidence: 99%

A prominent lack of IgG1-Fc fucosylation of platelet alloantibodies in pregnancy

Kapur¹,

Kustiawan

Vestrheim

et al. 2014

Blood

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285

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Intranasal Administration of Antibody-Bound Respiratory Syncytial Virus Particles Efficiently Primes Virus-Specific Immune Responses in Mice

Cited by 31 publications

References 47 publications

Multi-level glyco-engineering techniques to generate IgG with defined Fc-glycans

Multi-level glyco-engineering techniques to generate IgG with defined Fc-glycans

Glycosylation pattern of anti‐platelet IgG is stable during pregnancy and predicts clinical outcome in alloimmune thrombocytopenia

A prominent lack of IgG1-Fc fucosylation of platelet alloantibodies in pregnancy

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