OBJECTIVETo explore the suppression effect of human cytomegalovirus (HCMV) on megakaryocytes and their precursors and study the antiviral effect of antisense phosphorothioate deoxyoligonucleotide (ASON) against HCMV.METHODSCD34(+) cells were induced to proliferate and differentiate committedly to megakaryocytes in a semi-solid CFU-MK culture system. Cultured cells and ASON pretreated CD34(+) cells were infected by HCMV of AD169 strain. HCMV immediate early protein (IEP) DNA and mRNA and UL36 mRNA were detected by PCR and reverse transcription-polymerase chain reaction (RT-PCR). Cytotoxicity was evaluated by MTT assay.RESULTSHCMV AD169 suppressed the proliferation of megakaryocytes significantly. Compared with the mock group, the CFU-MK yields were decreased by 21.6%, 33.8%, and 46.3%, respectively, in 3 different titers of virus infected groups (P < 0.05). The suppression was virus titer dependent. HCMV IEP DNA, HCMV IEP mRNA and UL36 mRNA were detected in the colony cells of viral infection group. Compared with the infected group by HCMV AD169, UL36Anti treatment at 0.08 micromol/L could recover the CFU-MK yields significantly (P < 0.05). In the infected MK, which was pretreated with UL36Anti at 0.08 micromol/L, HCMV UL36 mRNA was undetectable by RT-PCR. The oligonucleotide MM(1) containing a G-to-C substitution in UL36Anti was inactive at 0.08 micromol/L but active at 0.40 micromol/L. The concentration of UL36Anti necessary to significantly affect cell growth was 90.00 micromol/L.CONCLUSIONSHCMV AD169 infection inhibits the proliferation and differentiation of megakaryocytes and their precursors. There are early transcriptions of HCMV IE and UL36 protein in infected CFU-MK. The specific ASON has a definite anti-HCMV activity.

01 May 1995·Antimicrobial Agents and ChemotherapyQ2 · MEDICINE

Potent antiviral activity of an antisense oligonucleotide complementary to the intron-exon boundary of human cytomegalovirus genes UL36 and UL37

Q2 · MEDICINE

Article

Author: Pari, G S ; Smith, J A ; Field, A K

An antisense phosphorothioate oligonucleotide complementary to the intron-exon boundary of human cytomegalovirus genes UL36 and UL37 (UL36ANTI) reduced the yield of infectious virus by 99% and inhibited human cytomegalovirus DNA replication at a concentration of 0.08 microM. In addition, oligonucleotides with base substitutions which resulted in base pair mismatches showed lesser degrees of activity, indicating a sequence-specific antisense mechanism. UL36ANTI was also shown to inhibit DNA replication of ganciclovir-resistant strains and human cytomegalovirus clinical isolates.

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Indication	Highest Phase	Country/Location	Organization	Date
Cytomegalovirus Infections	Preclinical	US	Idera Pharmaceuticals, Inc.	-

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