Highly passage of Spodoptera litura cell line causes its permissiveness to baculovirus infection

Zhang, Xuping; Lan, Wenjie; Deng, Yujie; Ma, Yuan; Liu, Kaiyu; Peng, Jianxin; Li, Yang; Hong, Hao

doi:10.1007/s10616-008-9158-8

Cited by 12 publications

(9 citation statements)

References 21 publications

(19 reference statements)

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“…7B). The titer of the budded baculovirus was about 2×10 8 pfu/ml for Sl-HP cells (Zhang et al 2008). But the titer of the supernatant from wild AcMNPV-infected HNU-Ha-MG1 cells was 4.2 ± 2 × 10 7 pfu/ml at 96 h post-infection.…”

Section: Chromosome Analysis Of Hnu-ha-mg1 Cellsmentioning

confidence: 92%

“…The osmolarity of the Grace's insect cell culture medium was about 300 mOsm, and the value of pH was 6.4. Spodoptera litura Sl-HP and T. ni Hi5 cells were cultured in Grace's insect cell culture medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin (Life Technologies, Inc., Carlsbad, CA) at 28°C (Zhang et al 2008). HEK293T cells were obtained from the American Tissue Culture Collection (Manassas, VA) and maintained in 5% CO 2 in DMEM supplemented with 10% fetal bovine serum (FBS).…”

Section: Methodsmentioning

confidence: 99%

“…The cells were observed under a fluorescence microscope at 24 h post-transfection, and the efficiency of the transfection was calculated according to the percentage of the cells emitting green fluorescence. To determine whether the cultured cells were contaminated by pathogens used in our laboratory such as Helicoverpa armigera densovirus and baculovirus, the cells cultured in flasks were also harvested by centrifugation and prepared for the observation of transmission electron microscope according to the method as described in our previous article (Zhang et al 2008). …”

Section: Methodsmentioning

confidence: 99%

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Establishment and characterization of a novel cell line from midgut tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)

Yang

et al. 2015

In Vitro Cell.Dev.Biol.-Animal

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The midgut of lepidopteran larvae serves as a target for many pathogens such as Bacillus thuringiensis (Bt). Cell lines originating from midgut tissues will be very helpful tools in many research fields. However, to date, no Bt-susceptible midgut-derived cell lines are available. Here, we reported that a novel cell line, designated as HNU-Ha-MG1, was established from midgut tissue of the fourth instar larvae of Helicoverpa armigera. This cell line grew well in Grace's insect cell culture medium supplemented with 10-15% fetal bovine serum. The shape of the most cells was round or polygonal, and some tended to aggregate to form multiple cell masses. The size of the cells was 13.8 ± 1.8 μm in diameter, and the maximum density reached (2.40 ± 0.15) × 10(6) cells/ml. The population doubling time during logarithmic growth phase was 58.6 ± 7.0 h at 28°C. The number of chromosomes was about 90-130, which exhibited typical chromosome characteristics of lepidopteran cell lines. The patterns of random amplified polymorphic DNA of the cell line were different from those of Sl-HP and Hi5 cell lines which were frequently used in our laboratory. 20-Hydroxyecdysterone induced apoptosis in a very small part of cells at 2 μg/ml but did not affect expression of autophagy-related protein 8 (Atg8) and its lipidation at 36 h post-treatment. The cell line was permissive to Autographa californica nuclear polyhedrosis virus (AcMNPV) and H. armigera nuclear polyhedrosis virus (HaSNPV). This cell line was found to be susceptible to activated Cry1C at the final concentration of 0.5-1.0 μg/ml but not to the activated Cry1Ac.

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Section: Chromosome Analysis Of Hnu-ha-mg1 Cellsmentioning

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Establishment and characterization of a novel cell line from midgut tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)

Yang

et al. 2015

In Vitro Cell.Dev.Biol.-Animal

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“…Spodoptera exigua larvae were raised on an artificial diet and reared at 25 ± 0.5°C under a 12L:12D photoperiod and 70% relative humidity. Sl‐HP cell line derived from Spodoptera litura ovaries was maintained in Grace's insect cell culture medium supplemented with 10% fetal bovine serum, 100 unit/ml penicillin, and 100 μg/ml streptomycin (Life Technologies Corporation) at 28°C (Zhang et al., ).…”

Section: Methodsmentioning

confidence: 99%

MOLECULAR CHARACTERIZATION OF AUTOPHAGY‐RELATED GENE 5 FROM Spodoptera exigua AND EXPRESSION ANALYSIS UNDER VARIOUS STRESS CONDITIONS

Liu

Xia

Zhou

et al. 2016

Arch Insect Biochem Physiol

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Autophagy is not only involved in development, but also has been proved to attend immune response against invading pathogens. Autophagy protein 5 (ATG5) is an important autophagic protein, which plays a crucial role in autophagosome elongation. Although ATG5 has been well studied in mammal, yeast, and Drosophila, little is known about ATG5 in lepidopteran insects. We cloned putative SeAtg5 gene from Spodoptera exigua larvae by the rapid amplification of cDNA ends method, and its characteristics and the influences of multiple exogenous factors on its expression levels were then investigated. The results showed that the putative S. exigua SeATG5 protein is highly homologous to other insect ATG5 proteins, which has a conserved Pfm domain and multiple phosphorylation sites. Next, fluorescence microscope observation showed that mCherry-SeATG5 was distributed in both nucleus and cytoplasm of Spodoptera litura Sl-HP cells and partially co-localized with BmATG6-GFP, but it almost has no significant co-localization with GFP-HaATG8. Then, the Western blot analysis demonstrated that GFP-SeATG5 conjugated with ATG12. Moreover, real-time PCR revealed that its expression levels significantly increased at the initiation of pupation and the stage of adult. In addition, the expression levels of SeAtg5 can be enhanced by the starvation, UV radiation, and infection of baculovirus and bacterium. However, the expression levels of SeAtg5 decreased at 24 h post treatments in all these treatments except in starvation. These results suggested that SeATG5 might be involved in response of S. exigua under various stress conditions.

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“…Spodoptera litura Sl‐HP, Sl‐1 (Zhang et al., ), Helicoverpa armigera QB‐Ha‐E‐5 (Ha) cell line (Zheng et al., ), B. mori BM‐21E‐HNU5 (Bme) cells (Li et al., ), and Lymantria dispar Ld652 cell lines (Goodwin et al., ) were grown in Grace's medium with 10% FBS and 1% penicillin‐streptomycin at 28°C.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Atg8 in Lepidopteran Insect Cells

Gai

Zhang

Islam

et al. 2013

Arch Insect Biochem Physiol

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Yeast Atg8 and mammalian microtubule-associated protein light chain 3 (LC3) are landmark proteins essential for autophagy. Here the lepidopteran Atg8, a homolog of LC3, is characterized. Sequence analysis reveals that Atg8 proteins are highly conserved in lepidopteran species. The abundance of endogeous Atg8 and the ratios of Atg8 conjugation to phosphatidylethanolamine (Atg8-PE)/Atg8 are different among several lepidopteran cell lines and different tissues of Helicoverpa armigera larvae. Both the density of fluorescent pre-autophagosomal structures with GFP-Ha Atg8 and the abundance of Atg6 are positively correlated with levels of Atg8-PE in different cell lines. The mutant GFP-Atg8(G116A) has lost the function in punctual formation, suggesting that G116 is important for autophagy. Exogenous factors have significant influences on the conversion of Atg8 in lepidopteran cells. Bacillus thuringiensis enhances the degradation of Atg8 in Spodoptera litura Sl-HP cells. Atg8-PE degrades gradually with extension of amino acid starvation, and bafilomycin A1 can block the decrease through the inhibition of autophagosome fusion with lysosome. Interestingly, high pH is more effective than amino acid starvation in Bombyx mori Bme cells to induce the conversion of BmAtg8 to BmAgt8-PE. Change of the quality of fetal bovine serum in the culture medium results in alteration of the ratio of Atg8-PE/Atg8 in some lepidopteran cell lines.

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Highly passage of Spodoptera litura cell line causes its permissiveness to baculovirus infection

Cited by 12 publications

References 21 publications

Establishment and characterization of a novel cell line from midgut tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)

Establishment and characterization of a novel cell line from midgut tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)

MOLECULAR CHARACTERIZATION OF AUTOPHAGY‐RELATED GENE 5 FROM Spodoptera exigua AND EXPRESSION ANALYSIS UNDER VARIOUS STRESS CONDITIONS

Characterization of Atg8 in Lepidopteran Insect Cells

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