Humanized antibodies reactive with L-selectin

Details Humanized antibodies reactive with L-selectin Humanized antibodies reactive with L-selectin

1995-12-27

2001-04-03

Gambel, Phillip (Department: 1644)

Drug, bio-affecting and body treating compositions

Immunoglobulin, antiserum, antibody, or antibody fragment,...

Monoclonal antibody or fragment thereof

C424S130100, C424S133100, C424S141100, C424S143100, C424S144100, C424S153100, C424S173100, C435S326000, C435S328000, C435S332000, C435S343000, C435S343100, C435S343200, C530S387100, C530S387300, C530S388100, C530S388200, C530S388220, C530S388700, C530S388730, C530S388750, C536S023100, C536S023500, C536S023530

Reexamination Certificate

active

06210671

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates generally to the combination of recombinant DNA and monoclonal antibody technologies for developing novel biologics and, more particularly, for example, to the production of non-immunogenic (in humans) immunoglobulins specific for the L-selectin protein and their uses in vitro and in vivo.
BACKGROUND OF THE INVENTION
The ability of cells to adhere to one another plays a critical role in development, normal physiology, and disease processes. This ability is mediated by adhesion molecules, generally glycoproteins, expressed on cell membranes. Often, an adhesion molecule on one cell type will bind to another adhesion molecule expressed on a different cell type, forming a receptor counter-receptor pair. Three very important classes of adhesion molecules are the integrins, selecting, and immunoglobulin (Ig) superfamily members (see Springer,
Nature
346:425 (1990); Osborn,
Cell
62:3 (1990); Hynes,
Cell
69:11 (1992), all of which are incorporated herein by reference in their entirety for all purposes). These molecules are especially vital to the interaction of leukocytes and platelets with themselves and with the extracellular matrix and vascular endothelium.
Integrins are heterodimeric transmembrane glycoproteins consisting of an &agr; chain (120-180 kD) and a &bgr; chain (90-110 kD), generally having short cytoplasmic domains. The &agr; subunits all share sequence homology and motifs with each other, as do the &bgr; subunits. The three known integrins containing the &bgr; subunit designated &bgr;
2
are important to the function of T cells, neutrophils and monocytes. LFA-1 (&agr;
L
&bgr;
2
) is widely distributed on lymphocytes, granulocytes and monocytes. Its counter-receptor is ICAM-1 (and perhaps of lesser importance, ICAM-2) an Ig family molecule which is expressed on many cells including leukocytes and is up-regulated on vascular endothelium by cytokines such as TNF and IL-1. Blocking LFA-1 on T cells with antibodies to either the &agr; or &bgr; subunit strongly inhibits adhesion-dependent functions such as CTL-mediated lysis of target cells. Mac-1 (&agr;
M
&bgr;
2
) is distributed on neutrophils and monocytes, and its counter-receptor is also ICAM-1 (and possibly ICAM-2). Among other things, Mac-1 is the type 3 complement receptor (CR3) and binds the C3bi fragment. The third &bgr;
2
integrin, P150,95 (&agr;
X
&bgr;
2
), is also found on neutrophils and monocytes, but seems of less importance. The a subunits of LFA-1, Mac-1 and P150,95 are also given the respective CD designations CD11a, CD11b and CD11c, while &bgr;
2
is also denoted CD18, so that LFA-1 is CD11a/CD18 and Mac-1 is CD11b/CD18.
There are three known selectins, which were previously known as LECCAMs, and are now designated L-selectin (also called LECAM-1, Mel-14 or LAM-1), E-selectin (also called ELAM-1) and P-selectin (also called GMP140 or PADGEM). They have all been sequenced at the cDNA level and share sequence homology and motifs, including a lectin-like domain. L-selectin has a dual role: it is a homing receptor on T cells for the high endothelial venules of peripheral lymph nodes, and it is an adhesion molecule on neutrophils for endothelium (Hallmann et al.,
Biochem. Biophys. Res. Commun.
174:236 (1991), which is incorporated herein by reference in its entirety for all purposes). E-selectin and P-selectin are both induced on endothelium by cytokines, although with different kinetics. L-selectin is a counter-receptor on neutrophils for both E-selectin and P-selectin (Picker et al.,
Cell
66:921 (1991), which is incorporated herein by reference in its entirety for all purposes), although all three selectins probably have other counter-receptors as well. In particular, E-selectin binds the carbohydrate group sialyl Lewis x (sLex) (Lowe et al.,
Cell
63:475 (1990)), which is incorporated herein by reference in its entirety for all purposes), and while this carbohydrate is prominently presented on L-selectin (Picker et al.,
Cell
66:921 (1991)), it may occur on other proteins as well. E-selectin is expressed especially in cutaneous sites of inflammation and also serves as an adhesion molecule for skin-homing T cells that may contribute to the inflammation (Picker et al.,
Nature
349:796 (1991), which is incorporated herein by reference in its entirety for all purposes).
In various assays, antibodies to CD11a, CD11b, CD18, L-selectin and E-selectin all block binding of neutrophils to activated endothelial cells to a lessor or greater degree, but the most complete inhibition is generally achieved by the combination of an antibody to CD18 and an antibody to L- or E-selectin (see, e.g., Luscinskas,
J. Immunol.
142:2257 (1989)), which is incorporated herein by reference in its entirety for all purposes). A recent but now widely accepted model accounts for these facts with a three step process of adhesion (Butcher,
Cell
67:1033 (1991), which is incorporated herein by reference in its entirety for all purposes). In the first step, neutrophils reversibly bind to inflamed vascular endothelium via the selecting, which bind well under conditions of flow, causing the neutrophils literally to roll along the vascular wall. The neutrophils are then activated by a variety of stimulants surrounding or released by the endothelium, including IL-8, PAF and C5a. The activated neutrophils shed L-selectin and up-regulate Mac-1. In the final step, binding of Mac-1 to ICAM-1 and perhaps other counter-receptors on the endothelial cells allows stable adhesion and extravasation through the endothelium.
In principle, antibodies or other antagonists of the integrin and selectin adhesion molecules could abort this process, by preventing neutrophils from binding to endothelium and from extravasating into tissues. Hence such antibodies could be used to treat a great many different disease conditions of which inflammation is an important component.
For example, in animal models anti-CD18 antibodies, which bind to both LFA-1 and Mac-1, have been useful in reducing ischemia-reperfusion injury (see, e.g., Vedder et al.,
J. Clin. Invest.
81:939 (1988); Vedder et al.,
Proc. Natl. Acad. Sci. USA
87:2643 (1990); U.S. Pat. No. 4,797,277). They also reduce neutrophil-mediated damage in the lung in response to various insults (Doerschuk et al.,
J. Immunol.
144:2327 (1990) and Mulligan et al.,
J. Immunol.
148:1847 (1992)), including gram-negative sepsis (Walsh et al.,
Surgery
110:205 (1991)). In a rabbit model, anti-CD18 antibodies also protect from lethality due to meningitis (Tuomanen et al.,
J. Exp. Med.
170:959 (1990)). They may also be useful in preventing or treating organ transplant rejection because-they block T-cell function.
For example, injection of antibodies to L-selectin or E-selectin into rodents suppressed neutrophil accumulation within inflamed peritoneum (Jutila et al.,
J. Immunol.
143:3318 (1989) and Mulligan et al.,
J. Clin. Invest.
88:1396 (1991)). Intravital video microscopy revealed that an anti-L-selectin antibody strongly inhibits rolling of leukocytes along the vascular wall endothelium of mesenteric venules exteriorized from rabbits (von Adrian et al.,
Proc. Natl. Acad. Sci. USA
88:7538 (1991)). An anti-E-selectin antibody greatly reduced vascular injury induced by immune complex deposition in the skin or lungs of rats, and substantially reduced neutrophil accumulation at those sites (Mulligen et al.,
J. Clin. Invest.
88:1396 (1991)). Also, in a primate model of extrinsic asthma, an anti-E-selectin antibody greatly reduced neutrophil influx into the lung and associated late-phase airway obstruction after antigen inhalation (Gundel et al.,
J. Clin. Invest.
88:1407 (1991)).
Several antibodies including mouse DREG-55, mouse DREG-56 and mouse DREG-200 have been developed that bind to human L-selectin (Kishimoto et al.,
Proc. Natl. Acad. Sci. USA
87:2244 (1990), which is incorporated herein by reference in its entirety for all purposes). These antibodies partially or completely block the binding of human lymphocytes to peripheral lymph nod

Affiliated with

Also associated with

Rating

Humanized antibodies reactive with L-selectin does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Humanized antibodies reactive with L-selectin, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Humanized antibodies reactive with L-selectin will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2435084