PMID- 35405136
OWN - NLM
STAT- MEDLINE
DCOM- 20220603
LR  - 20220613
IS  - 1096-0805 (Electronic)
IS  - 0022-2011 (Linking)
VI  - 191
DP  - 2022 Jun
TI  - Heterologous expression of scFv fragment against Vairimorpha (Nosema) ceranae 
      hexokinase in Sf9 cell culture inhibits microsporidia intracellular growth.
PG  - 107755
LID - S0022-2011(22)00040-4 [pii]
LID - 10.1016/j.jip.2022.107755 [doi]
AB  - Secretion of hexokinase (HK) by microsporidia into infected cells suggests an 
      important role for this enzyme for the intracellular development of parasites. To 
      verify whether the expression of HK-specific antibodies in the host cell 
      cytoplasm can suppress the growth of microsporidia, we constructed an immune 
      library of recombinant scFv fragments against the enzyme of the honey bee 
      pathogen Vairimorpha (Nosema) ceranae (VcHK) with a representativeness of about 5 
      million bacterial transformants. Two variants of VcHK-specific recombinant 
      antibodies were selected by library panning and expressed in lepidopteran Sf9 
      cell line. Infecting of cells expressing two selected and control scFv fragments 
      with V. ceranae spores was followed by their cultivation for 4 days. Analysis of 
      parasite beta-tubulin as well as spore wall protein SWP32 transcripts in infected 
      cultures by reverse transcription PCR and real-time qPCR showed (1) V. ceranae 
      growth in cells heterologous to bee pathogens, (2) its inhibition by one of the 
      selected VcHK-specific recombinant antibodies. The latter result once again 
      emphasizes an important role of microsporidia hexokinases in their relationships 
      with infected host cells and suggests further focusing on the mechanisms of such 
      suppression, as well as on the search for new V. ceranae - inhibiting scFv 
      fragments.
CI  - Copyright (c) 2022 Elsevier Inc. All rights reserved.
FAU - Dolgikh, Viacheslav V
AU  - Dolgikh VV
AD  - Laboratory of Molecular Plant Protection, All-Russian Institute of Plant 
      Protection, St. Petersburg, Pushkin, Russia. Electronic address: 
      dol1slav@yahoo.com.
FAU - Zhuravlyov, Vladimir S
AU  - Zhuravlyov VS
AD  - Laboratory of Molecular Plant Protection, All-Russian Institute of Plant 
      Protection, St. Petersburg, Pushkin, Russia.
FAU - Senderskiy, Igor V
AU  - Senderskiy IV
AD  - Laboratory of Molecular Plant Protection, All-Russian Institute of Plant 
      Protection, St. Petersburg, Pushkin, Russia.
FAU - Ignatieva, Anastasiya N
AU  - Ignatieva AN
AD  - Laboratory of Molecular Plant Protection, All-Russian Institute of Plant 
      Protection, St. Petersburg, Pushkin, Russia.
FAU - Timofeev, Sergey A
AU  - Timofeev SA
AD  - Laboratory of Molecular Plant Protection, All-Russian Institute of Plant 
      Protection, St. Petersburg, Pushkin, Russia.
FAU - Seliverstova, Elena V
AU  - Seliverstova EV
AD  - Laboratory of Renal Physiology, Sechenov Institute of Evolutionary Physiology and 
      Biochemistry, St. Petersburg, Russia.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220409
PL  - United States
TA  - J Invertebr Pathol
JT  - Journal of invertebrate pathology
JID - 0014067
RN  - EC 2.7.1.1 (Hexokinase)
RN  - Nosema ceranae
RN  - Vairimorpha cheracis
SB  - IM
MH  - Animals
MH  - Bees
MH  - Cell Culture Techniques
MH  - Hexokinase
MH  - Microsporidia
MH  - *Nosema/physiology
OTO - NOTNLM
OT  - Hexokinase
OT  - Honey bee
OT  - Microsporidia
OT  - Single-chain antibodies
OT  - Vairimorpha (Nosema) ceranae
OT  - scFv immune library
EDAT- 2022/04/12 06:00
MHDA- 2022/06/07 06:00
CRDT- 2022/04/11 20:10
PHST- 2021/10/19 00:00 [received]
PHST- 2022/03/31 00:00 [revised]
PHST- 2022/04/05 00:00 [accepted]
PHST- 2022/04/12 06:00 [pubmed]
PHST- 2022/06/07 06:00 [medline]
PHST- 2022/04/11 20:10 [entrez]
AID - S0022-2011(22)00040-4 [pii]
AID - 10.1016/j.jip.2022.107755 [doi]
PST - ppublish
SO  - J Invertebr Pathol. 2022 Jun;191:107755. doi: 10.1016/j.jip.2022.107755. Epub 
      2022 Apr 9.