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Interleukin 4 enhances osteoblast macrophage colony-stimulating factor, but not interleukin 6, production (1994)

Lacey, D. L. ; Erdmann, J. M. ; Shima, M. ; [et al.]

Springer

Calcified tissue international 55 (1994), S. 21-28

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ISSN: 1432-0827

Keywords: M-CSF mRNA ; IL-1-B9 cells ; 5A1 ; M-CSF ELISA

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine , Physics

Notes: Abstract To determine if interleukin 4's (IL-4) recently discovered skeletal effects could be explained by its effects on osteoblasts, we have examined IL-4's impact on macrophage colony stimulating factor (M-CSF) and interleukin 6 (IL-6) secretion by the murine osteoblastic cell line MC3T3-E1. Interleukin-4 increased colony-forming activity in MC3T3 supernatants two-threefold with colony cytomorphology, cytohistochemistry, and blockade of the effect by anti-M-CSF antibody, indicating that the IL-4-induced activity was M-CSF. MC3T3 M-CSF supernatant activity increased in a time-dependent manner with positive IL-4 effects seen after a 24-hour exposure. The maximal IL-4 effective dose was 100 U/ml where conditioned media from IL-4-treated cells contained twofold more M-CSF than control cells (400 U/ml versus 200 U/ml M-CSF) as detected by a sandwich M-CSF ELISA. Northern blots showed that IL-4 (200 U/ml) rapidly increased steady-state M-CSF mRNA levels with maximal induction observed by 2 hours followed by a decline to near basal levels by 24 hours. IL-4 also dose dependently increased M-CSF mRNA levels with maximal induction (fourfold) seen at 100 U/ml IL-4. In contrast to its impact on MC3T3 M-CSF production, IL-4 (200 U/ml) did not stimulate MC3T3 IL-6 secretion whereas IL-1 (1 pM) stimulated a 500-fold increase in MC3T3 IL-6 release. When utilized to treat newborn calvarial osteoblast-enriched cultures, IL-4 dose dependently augmented M-CSF production, with the maximal effect seen at 200 U/ml where IL-4-treated, osteoblast-conditioned media contained almost 500 U/ml M-CSF, compared with 200 U/ml M-CSF in controlconditioned media. These observations indicate that the range of IL-4 cellular targets in skeletal tissues include osteoblastic cells, and that this cytokine increases osteoblast expression of M-CSF, a hematopoietic cytokine pivotal for monocyte/macrophage differentiation. Furthermore, IL-4's impact on osteoblast-produced M-CSF levels is selective because IL-6 levels were unaltered by IL-4 treatment.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00310164

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Interleukin 4 increases type 5 acid phosphatase mrna expression in murine bone marrow macrophages (1994)

Lacey, D. L. ; Erdmann, J. M. ; Tan, H.-L.

New York, N.Y. : Wiley-Blackwell

Journal of Cellular Biochemistry 54 (1994), S. 365-371

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ISSN: 0730-2312

Keywords: vitamin D ; glucocorticoids ; polymerase chain reaction ; osteoclasts ; M-CSF Abbreviations: TRAP ; tartrate resistant acid phosphatase; RT-PCR ; reverse transcription-polymerase chain reaction; IL-4 ; interleukin 4 ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Chemistry and Pharmacology , Medicine

Notes: Type 5 acid phosphatase is a lysosomal enzyme expressed in cells of monocyte/macrophage lineage frequently used as a marker of osteoclastic differentiation. Oligonucleotide primers for DNA amplification were designed following sequence alignment of rat bone and human macrophage type 5 acid phosphatases. DNA (330 bp in length) obtained using these primers and reverse transcribed total cell RNA from in vitro generated murine osteoclastic cells was cloned and sequenced. DNA sequence analysis of two clones demonstrates that the amplified material was 91% and 96% identical to rat bone type 5 acid phosphatase at the nucleotide and amino acid level, respectively. Northern blots of murine tissue RNA show the presence of 1.5-kb transcripts that are most highly expressed in the long bones. Total cell RNA from the osteoclastic cells contain a marked level of type 5 acid phosphatase mRNA when compared to the levels seen in the tissue samples. Additionally, osteoclastic cell RNA contains two additional transcripts of 2.5 and 5 kb. Bone marrow macrophages grown in the presence of M-CSF express low levels of the 1.5-kb transcript with no signal observed for either of the two larger transcripts that were seen in the osteoclastic RNA samples. Importantly, bone marrow macrophage 1.5-kb type 5 acid phosphatase transcript levels are increased by interleukin 4 treatment in both a time and concentration-dependent manner. These findings indicate that type 5 acid phosphatase, while a cytochemical marker for osteoclasts, can be induced in macrophages by agents that block in vitro osteoclastic differentiation. Increased type 5 acid phosphatase may play a role in interleukin 4-stimulated monocyte activities.

Additional Material: 5 Ill.