Cloning and expression of bovine and porcine interleukin-2 in baculovirus and analysis of species cross-reactivity

Collins, Robert A.; Tayton, H.K.; Gelder, K.I.; Britton, Paul; Oldham, G.

doi:10.1016/0165-2427(94)90042-6

Cited by 37 publications

(23 citation statements)

References 21 publications

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“…Supernatants from cells infected with the recombinant BHV-1 constructs yielded comparable levels of bioactive boiL-2 and boiL-4, respectively, and antibodies which neutralized cytokine function provided a convenient way to confirm the specificity of these assays. In agreement with previous studies on the bioactivity of rbo-IL-2, BHV-I-encoded boiL-2 induced the proliferation of activated B (Collins & Oldham, 1995) and T (Collins et al, 1994) cells. This contrasted with the findings with boiL-4, which induced B cell proliferation, but was not capable of inducing T cell proliferation by itself, although it enhanced boIL-2-driven proliferation (data not shown).…”

Section: Discussionsupporting

confidence: 92%

“…Because plasmid pACIL-2.B222 (Collins et al, 1994) encompasses only codons 21-155 of the boiL-2 gene (Cerretti et al, 1986), the boiL-20RF was completed using synthetic oligonucleotides and standard cloning procedures (Sambrook et al, 1989). The sequence contained in the resulting plasmid, pSPIL-2OK, was verified by dideoxy chain-termination sequencing (T7 sequencing kit; Pharmacia).…”

Section: Methodsmentioning

confidence: 99%

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Bovine Interleukins 2 and 4 Expressed in Recombinant Bovine Herpesvirus 1 are Biologically Active Secreted Glycoproteins

Kühnle

Collins

Scott

et al. 1996

Journal of General Virology

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Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Bovine Interleukins 2 and 4 Expressed in Recombinant Bovine Herpesvirus 1 are Biologically Active Secreted Glycoproteins

Kühnle

Collins

Scott

et al. 1996

Journal of General Virology

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“…In contrast to bovine T cells that proliferate in response to human recombinant IL-2 [28], bovine IL-2 was required for the culture of bovine NK cells. Bovine NKp46 + cells could also to some extent be expanded in supernatant from Con A-stimulated bovine CD4 + T cells (data not shown).…”

Section: (D) Nkp46mentioning

confidence: 99%

NKp46 defines a subset of bovine leukocytes with natural killer cell characteristics

et al. 2004

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Natural killer (NK) cells have not previously been precisely identified or characterized in cattle or any other ruminant species. We have generated a monoclonal antibody against bovine NKp46, which is expressed exclusively by NK cells in man. NKp46 + cells comprised 1-10% of blood mononuclear cells in cattle, and did not stain with antibodies against CD3, CD4, TCR1, B cell or granulocyte markers. The majority of the NKp46 + cells expressed CD2, and a variable fraction also expressed CD8. The tissue distribution of NKp46 + cells in cattle was compatible with the tissue distribution of NK cells in other species. Bovine NKp46 + cells had typical, large granular lymphocyte morphology, and proliferated vigorously in response to bovine IL-2 for a limited number of cell divisions. IL-2-activated NKp46 + cells killed the bovine kidney cell line MDBK. This cytotoxicity was inhibited by preincubation with antibody against NKp46. In a redirected lysis assay, IL-2-activated NKp46 + cells killed the Fc + R + target cell line P815 after preincubation with antibody against NKp46. Together, these data indicate that bovine NKp46 is an activating receptor and demonstrate the existence of a subset of leukocytes in cattle that, in terms of surface markers, morphology and function, represent NK cells.

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“…1B. It was considered that the band with 17 kDa was a precursor form of rbIL-2 with signal peptide and the other band was a mature form of rbIL-2 as reported previously [8].…”

mentioning

confidence: 92%

Expression of Bovine Interleukin-2 in Insect Cells by Recombinant Baculovirus

Kashima¹,

Iwata²,

Inoue³

1999

The Journal of Veterinary Medical Science

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ABSTRACT. Bovine interleukin-2 (bIL-2) was expressed in insect cells infected with recombinant baculovirus. The cDNA for bIL-2 was amplified by reverse transcriptional polymerase chain reaction and recombinant baculovirus was constructed by homologous recombination. The recombinant baculovirus was plaque-purified, amplified, and then infected to Sf9 cells for expression of recombinant bIL-2 (rbIL-2). In result, the protein band corresponding to rbIL-2 could be detected on SDS-PAGE with coomassie brilliant blue staining and on immunoblot analysis reacted with mouse antiserum against bIL-2. In addition, cell proliferation assay for bIL-2 activity demonstrated that the culture supernatant of Sf9 cells infected with recombinant baculovirus enhanced the proliferation of bovine peripheral blood mononuclear cells in a dose-dependent manner.-KEY WORDS: bovine interleukin-2, insect cell, recombinant baculovirus

show abstract

Cloning and expression of bovine and porcine interleukin-2 in baculovirus and analysis of species cross-reactivity

Cited by 37 publications

References 21 publications

Bovine Interleukins 2 and 4 Expressed in Recombinant Bovine Herpesvirus 1 are Biologically Active Secreted Glycoproteins

Bovine Interleukins 2 and 4 Expressed in Recombinant Bovine Herpesvirus 1 are Biologically Active Secreted Glycoproteins

NKp46 defines a subset of bovine leukocytes with natural killer cell characteristics

Expression of Bovine Interleukin-2 in Insect Cells by Recombinant Baculovirus

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