Salama, A. | Search bioRxiv

bioRxiv2020

High neutralizing potency of swine glyco-humanized polyclonal antibodies against SARS-CoV-2

Perfusion of convalescent plasma (CP) has demonstrated a potential to improve the pneumonia induced by SARS-CoV-2, but procurement and standardization of CP are barriers to its wide usage. Many monoclonal antibodies (mAbs) have been developed but appear insufficient to neutralize SARS-CoV-2 unless two or three of them are being combined. Therefore, heterologous polyclonal antibodies of animal origin, that have been used for decades to fight against infectious agents might represent a highly efficient alternative to the use of CP or mAbs in COVID-19 by targeting multiple antigen epitopes. However, conventional heterologous polyclonal antibodies trigger human natural xenogeneic antibody responses particularly directed against animal-type carbohydrate epitopes, mainly the N-glycolyl form of the neuraminic acid (Neu5Gc) and the Gal 1,3-galactose (Gal), ultimately forming immune complexes and potentially leading to serum sickness or allergy. To circumvent these drawbacks, we engineered animals lacking the genes coding for the cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) and 1,3-galactosyl-transferase (GGTA1) enzymes to produce glyco-humanized polyclonal antibodies (GH-pAb) lacking Neu5Gc and -Gal epitopes. We found that pig IgG Fc domains fail to interact with human Fc receptors and thereby should confer the safety advantage to avoiding macrophage dependent exacerbated inflammatory responses, a drawback possibly associated with antibody responses against SARS-CoV-2 or to avoiding a possible antibody-dependent enhancement (ADE). Therefore, we immunized CMAH/GGTA1 double knockout (DKO) pigs with the SARS-CoV-2 spike receptor-binding domain (RBD) to elicit neutralizing antibodies. Animals rapidly developed a hyperimmune response with anti-SARS-CoV-2 end-titers binding dilutions over one to a million and end-titers neutralizing dilutions of 1:10,000. The IgG fraction purified and formulated following clinical Good Manufacturing Practices, named XAV-19, neutralized Spike/angiotensin converting enzyme-2 (ACE-2) interaction at a concentration < 1g/mL and inhibited infection of human cells by SARS-CoV-2 in cytopathic assays. These data and the accumulating safety advantages of using glyco-humanized swine antibodies in humans warranted clinical assessment of XAV-19 to fight against COVID-19.

immunology↗

bioRxiv2019

Prenatal heat stress effects on gestation and postnatal behavior in kid goats

Consequences of heat stress during pregnancy can affect the normal development of the offspring. In the present experiment, 30 Murciano-Granadina dairy goats (41.8 {+/-} 5.7 kg) were exposed to 2 thermal environments varying in temperature-humidity index (THI) from 12 days before mating to 45 days of gestation. The environmental conditions were: gestation thermal-neutral (GTN; THI = 71 {+/-} 3); and gestation heat stress (GHS; THI = 85 {+/-} 3). At 27 {+/-} 4 days old, GTN-born female kids (n = 16) and GHS-born ones (n = 10) were subjected to 2 tests: arena test (AT) and novel object test (NOT), the latter was repeated at 3 months of age. Additionally, 8 months after birth, a subset of growing goats (n = 8) coming from GTN and GHS (16.8 {+/-} 3.4 kg BW) were exposed consecutively to 2 environmental conditions: a basal thermal-neutral period (THI = 72 {+/-} 3) for 7 days, and a heat-stress period (THI = 87 {+/-} 2) for 21 days. In both periods, feeding behavior, resting behavior, other active behaviors (exploring, grooming), thermally-associated behaviors and posture were recorded. The gestation length was shortened by 3 days in GHS goats. In the AT, GHS kids showed a lower number of sniffs (P < 0.01) compared to GTN. In the NOT, GHS kids also tended to show a lower number of sniffs (P = 0.09). During heat exposure, GTN and GHS growing goats spent more time resting as well as exhibited more heat-stress related behaviors such as panting and drinking (P < 0.001); however, no differences were observed between both groups. In conclusion, heat stress during the first third of pregnancy shortened gestation length and influenced the exploratory behavior of the kids in the early life without impact on the behavior during the adulthood when exposed to heat stress.

animal behavior and cognition↗