A monoclonal antibody against an antigen present on mouse macrophages and absent from monocytes

Abstract

A monoclonal antibody (BM8) raised in the rat against cultured mouse bone marrow-derived macrophages reacted only with macrophages and not with granulocytes, mast cells, platelets, lymphocytes, fibroblasts, and endothelial cells. BM8 did not detect blood monocytes. In cultured bone-marrow cells, expression of BM8-antigen was found on a few macrophages after one day of culture and reached its maximum level with 80% positive macrophages after 7-10 days of culture. The antibody BM8 belonged to the IgG2a subclass, was non-cytotoxic and directed against a 125 kD membrane antigen. In cryostat sections of normal mouse tissues (spleen, lymph node, thymus, liver, skin) BM8 detected tissue-fixed macrophages and Langerhans cells in the skin. In spleen and lymph node, BM8 reacted with macrophages in the red pulp and in the medullary cords, respectively, but not with heavily phagocytosing marginal-zone macrophages, as revealed by in-vivo phagocytosis of colloidal carbon. In granulomata induced by complete Freund's adjuvant, BM8 detected inflammatory macrophages but not epithelioid cells. Thus, antibody BM8 detected a differentiation antigen expressed only on mature, tissue-fixed macrophages.

Smad3 Knockout Mice Exhibit a Resistance to Skin Chemical Carcinogenesis

“…To examine inflammatory cell subtypes, immunohistochemistry was performed on frozen sections, as we have described previously (16) (26), was also used. Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and phosphorylated Smad2 was performed on paraffin-embedded sections using a PCNA antibody (1:100; Santa Cruz Biotechnology) and a phosphorylated Smad2 antibody (1:50; Cell Signaling, Beverly, MA).…”

Retinal degeneration triggered by inactivation of PTEN in the retinal pigment epithelium

“…The overproduction of MG in the mutant retina is common to many retinal degenerative diseases (Bringmann and Reichenbach 2001), and is normally induced by the inflammation that is triggered by infiltrating macrophages and monocytes. We monitored the distribution of macrophages by immunostaining with the pan-macrophage marker F8/40 (Malorny et al 1986). F8/ 40-positive macrophages were confined outside of the RPE layer in wild-type mouse eyes, but were detected in the subretinal spaces and the inner plexiform layer (IPL) of fl/fl;T1Cre mouse retinae (data not shown).…”

Two Different Types of Sialoadenitis in the NOD- and MRL/lpr Mouse Models for Sjögren's Syndrome: A Differential Role for Dendritic Cells in the Initiation of Sialoadenitis?

“…This was then added to 165 ml of distilled water in which the slides were fixed. After the fixation, the slides were rinsed in phosphatebuffered saline (PBS) (pH 7.8)/Tween (0.1%), after which they were incubated for 1 hour with the monoclonal antibody N418 (Metlay et al, 1990), KT3 (Tomonari, 1988), B220 (Ledbetter and Herzenberg, 1979), or BM8 (Malorny et al, 1986). Except for BM8 (BMA, Biomedical AG, Augst, Switzerland), all monoclonal antibodies were hybridoma culture supernatants reacting with DC, total T cells, and B cells, respectively.…”