BACTERICIDAL PERMEABILITY INCREASING PROTEIN (BPI) FOR TREATING CONDITIONS ASSOCIATED WITH CORNEAL INJURY

Abstract: The present invention provides methods of treating a subject suffering from adverse effects, complications or conditions, associated with or resulting from a corneal injury including, corneal infection or ulceration, by topical administration of suitable ophthalmic preparations of bactericidal/permeability-increasing (BPI) protein products.

Description: BACTERICIDAL PERMEABILITY INCREASING PROTEIN (BPI) FOR TREATING CONDITIONS ASSOCIATED WITH CORNEAL INJURY. 5 BACKGROUND OF THE INVENTION The present invention relates generally to methods of treating a subject suffering from adverse effects, complications or conditions including infection or ulceration associated with or resulting from corneal injury from, for example, perforation, abrasion, chemical burn or trauma injury, by topical 10 administration of bactericidal/permeability-increasing (BPI) protein products. Corneal infections, microbial keratitis and infectious comeal ulceration are increasingly prevalent, serious and sight-threatening ophthalmic diseases. Infectious or microbial keratitis is an infection of the cornea characterized by an ulceration of the co eal epithelium associated with an 15 underlying inflammatory infiltrate of the comeal stroma. Infectious keratitis is the most serious complication of wearing contact lenses. Complications of infectious keratitis include sight-threatening scar formation, scleral involvement, comeal perforation, and even loss of the eye. Comeal diseases are estimated to involve several hundredthousand cases of corneal ulcers and 20 about twice that number of keratitis cases each year in the U.S. alone. Contact lens wearers, immunocompromised individuals and patients suffering from dry eye syndrome are among those most at risk to develop such co eal lesions. In third world countries, this cause of blindness is second only to cataract formation. 25 Microbial keratitis, or infections of the cornea, can be caused by various bacteria, fungi, viruses, or parasites. Bacteria are the most common causes, but the frequency of involvement of different species may vary from one geographic region to another and may show a shifting pattern over time. Species of bacteria causing keratitis in the majority of cases are: 30 (1) Micrococcaceae (Staphylococcus, Micrococcus), (2) Streptococci, (3) Pseudomonas, and (4) Enterobacteriaceae (Citrobacter, Klebsiella, Enterobacter, Serratia, Proteus). Historically, the pneumococcus (Streptococcus pneumoniae) was a major cause, but now other gram-positive organisms predominate, with Staphylococcus aureus reported to be the most common cause of microbial keratitis in the northern United States. Pseudomonas aeruginosa has also become more prevalent as a cause of keratitis, particularly in association with overnight contact lens wear. Infections involving the indigenous bacteria of the conjunctiva and eyelids (Staphylococcus epidermidis, Corγnebacterium and Propionibacterium species) are reportedly being seen more frequently, as are other commensal and less virulent organisms, especially in immunocompromised hosts. The variety of organisms most commonly seen in bacterial keratitis has been documented (see, e.g., Liesegang, Bacterial Keratitis, in Infectious Disease Clinics of North America, Vol. 6, No. 4, pp.815-829, December, 1992); however, any organism, under appropriate circumstances, can be a causative agent of comeal infection and ulceration. Comeal infection is usually precipitated by an epithelial defect resulting from injury (including perforation, abrasion, chemical bu or trauma injury) to the cornea or from contact lens wear. Co eal disease patients and patients receiving topical corticosteroids or with compromised local or systemic defense mechanisms appear more susceptible to co eal epithelial defects precipitating infection. The cornea is an avascular structure, and has a protective coating with two layers of mucosubstances, including an adherent glycocalyx and a mucin layer produced by goblet cells. The intact comeal epithelium is usually an effective barrier against infection, although some bacterial organisms, notably Neisseria gonorrhoeae and Corynebacterium diphtheriae, can penetrate the intact epithelium. The lids and eyelashes normally harbor microorganisms and shed them onto the cornea, but the eyelids provide a defensive system for the co ea, primarily through the lacrimal secretions and the ocular blink reflex. The tear film provides lubrication to flush away any organisms or debris. The tear film also contains several antimicrobial substances, including lysozyme, lactofeπϊn, beta-lysins, and complement components, as well as immunoglobulins (especially secretory IgA) and lymphocytes, which provide a local defense mechanism. Lactoferrin can enhance the effect of surface antibodies or inhibit bacterial growth or invasiveness by chelating iron. Tear lysozyme can directly lyse bacterial cell walls, and beta-lysins can lyse bacterial membranes. Secretory IgA blocks the adhesion of bacteria to membranes. Malposition of the lids and lashes, however, or difficulty in lid closure interferes with these protective functions and predisposes to co eal infection. Predisposing factors to comeal infection therefore include: (1) trauma or injury (e.g., foreign body, contact lens wear); (2) abnormal tear function (e.g., dry eye, lacrimal obstruction) and abnormal lid structure and function (e.g., blepharitis, laopthalmus entropion, ectropion, trichiasis); (3) comeal diseases (e.g., comeal edema); and (4) systemic conditions (e.g. , Sjόgren's syndrome, alcoholism, diabetes, rheumatoid arthritis, debilitating disease, tracheal intubation, central nervous system disease and psychiatric disturbances, extensive bums, acquired immunodeficiency syndrome (AIDS), and corticosteroid and immunosuppressive therapy). Contact lens wear is a significant risk factor compromising the structural integrity of the comeal epithelium and predisposing toward co eal infection. Contact lens wear give rise to comeal hypoxia, increased comeal temperature, decreased tear flow to the cornea, and also provides a constant source of microtrauma to the co eal epithelium. Soft contact lenses become coated with mucus and protein after only a few hours of wear, and this may further enhance the adherence of bacteria. Hard gas-permeable lenses, daily wear soft contact lenses, extended wear soft contact lenses, therapeutic soft contact lenses, and disposable contact lenses all increase the risk of microbial keratitis. Overnight wear, especially after cataract surgery, is associated with the highest risk. Other factors contributing to contact lens-associated microbial keratitis include the failure to follow proper contact lens wear instructions, poor contact lens hygiene, use of contaminated lens solutions, and microtrauma at the time of the insertion and removal. Pseudomonas aeruginosa and Staphylococcus are the most common organisms isolated in contact lens-associated keratitis. Acanthamoeba keratitis, a parasitic infection, has been linked to prolonged exposure to contaminated water, especially in contact lens wearers and in individuals who use hot tubs or swimming pools. Fungal keratitis is seen in different clinical situations. Filamentary fungal keratitis is seen after injury to the cornea in agricultural settings, whereas yeast keratitis is seen in any environment in patients who are immunocompromised or have a severely damaged cornea. The severity of the bacterial keratitis depends, for the most part, on the virulence of the invading bacteria but also is correlated to the previous health of the cornea and the host response. The pathogenicity of particular organisms is correlated with the ability to adhere to the edge or base of an epithelial defect and to invade the comeal stroma. Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pneumoniae adhere tightly to the edge of an epithelial defects, probably because of membrane appendages called fibrillae (in gram-positive organisms) or fimbriae (in gram- negative organisms). Specific adhesions on the surface of these appendages may interact with specific receptors on the comeal epithelium. Some species, notably Pseudomonas and Staphylococcus, produce an extracellular polysaccharide slime layer which may have a role in adherence to a variety of surfaces, especially soft contact lenses. The mechanisms of penetration of bacteria into the co eal stroma following entry through an epithelial injury are poorly understood but are probably correlated with the production of toxins and enzymes. Pseudomonas and Serratia species have proteoglycanase (e.g. , collagenase) activity that can liquify the stroma. Other organisms have other properties that permit adherence and comeal destruction. The host's polymorphonuclear response to the infection contributes to the tissue destruction and collagen breakdown as a result of lysozymal enzymes and other proteases. In a previously healthy cornea, the presence of a co eal epithelial ulceration with adherent mucopurulent exudate and inflammatory cells in the adjacent co eal stroma and the anterior chamber should lead to a presumptive diagnosis of bacterial keratitis. The eyelids may be stuck together and the tear film filled with inflammatory cells. Nonspecific symptoms include decreased vision, redness, pain, conjunctival and lid swelling and a discharge. Clinical signs may include increasing stromal edema, hypopyon, iris miosis, and synechiae. In a patient with a cornea previously damaged by herpes simplex virus infection, comeal edema, or trauma, it may be difficult to distinguish the clinical signs of infection from the residua of the underlying structural abnormalities. A bacterial infection should be suspected when there is an increase in the extent of epithelial or stromal ulceration or anterior chamber inflammation. Antecedent therapy with systemic or local ocular immunosuppressive agents, especially corticosteroids, not only increases the risk of ocular infection but may alter the clinical response in such a way as to mask or alter some of the typical features of infection. There are difficulties in distinguishing bacterial keratitis from other forms of microbial keratitis or from the multiple noninfectious causes of comeal ulceration. The differential diagnosis includes fungal, viral, and parasitic keratitis as well as toxic or chemical keratopathy, indolent or neurotrophic ulceration, severe dry eyes, and various other insults to the cornea. The history, physical examination, and evidence of the onset of the new disease process may permit a presumptive diagnosis. When comeal infection is suspected, the culture strategy may include screening for the most likely agents: aerobic bacteria, anaerobic bacteria, filamentous fungi, and yeasts. A co eal sample may be obtained by scraping, using the magnification of the slit lamp biomicroscope, and topical anesthesia. With deep keratitis, fragments of the cornea may be excised with a microsurgical scissor or trephine. More than one species of microbe may be present in a comeal infection. Negative cultures are not uncommon in cases of suspected infectious comeal ulcers, and may be due to inadequate sampling methods, the improper selection of media, prior antibiotic treatment, or improper interpretation of data. Currently, the initial therapy for suspected microbial keratitis is based on the severity of the keratitis and a familiarity with the most likely causative organisms. Suspected microbial keratitis is typically treated as a bacterial ulcer until a more definitive laboratory diagnosis is made. Initial antibiotic therapy may be based on the results of the Gram stain or Giemsa stain, or a broad spectrum antibiotic may be administered as the initial treatment, especially in cases of serious suspected microbial keratitis. Most U.S. practitioners are not willing to leave the lesion untreated while waiting for culture results. Generally, a broad spectrum antibiotic is prescribed following examination. Such initial antibiotic therapy may be modified after the causative organism is identified from correlation of the Gram stain, culture results, and the clinical response. There are a relatively small number of antibiotics available commercially as topical ophthalmic preparations. Many other antibiotics can be prepared for topical ophthalmic use in treating serious co eal infections, however, their use is expensive and inconvenient, and many are not well tolerated or have limited antibacterial spectra. Pseudomonas species account for many serious, and rapidly destructive, comeal infections. In fact, ocular disease produced by the opportunistic bacterial pathogen P. aeruginosa often leads to a fulminating and highly destructive infection resulting in rapid liquefaction of the cornea and blindness. Antibiotic treatment is not always successful due to the resistance of many clinical strains. The patient is vulnerable during the ulcerative period to sequelae that are sight threatening and even could create a situation where the eye had to be enucleated. Any agent that could accelerate the healing time, for example, would be highly desired by medical practitioners. Thus, there is an unmet need to develop agents with therapeutic efficacy, either alone or in conjunction with existing agents, against these organisms. In cases where there is the need for frequent administration of antimicrobial drops and the need to examine the patient daily, patients may be hospitalized. Patient isolation is not usually necessary, although contact with preoperative patients should be avoided. Outpatient therapy may be preferred for compliant patients or those with milder disease. The ideal topical antibiotic agent should be bactericidal at reasonable concentrations against the comeal pathogens, should be able to penetrate the cornea, and should be free of significant adverse affects. Factors considered in the use of systemic antibiotics (i.e., achievable serum levels, distribution space, and absorption and excretion characteristics) are not applicable. Some patients may respond to commercial-strength topical antibiotic agents given at frequent intervals, but fortified topical antibiotic agents are usually more effective. For example, recent fluoroquinolone antibiotics, norfloxacin and ciprofloxacin, may be effective at commercial strength for infections by susceptible bacteria. Drug penetration into the cornea may be increased with higher concentration of the drug, more frequent application, longer contact time with the use of some vehicles, with more lipophilic antibiotic agents, and with the absence of the epithelium. Solutions may be preferred to ointments because of the flexibility in varying the concentration and the ease of administration. A fortified topical antibiotic agent may be prepared by adding the desired amount of the parenteral antibiotic to an artificial tear solution. The primary goal of current therapy is to administer an antibiotic which will be effective quickly without causing significant ocular and systemic toxicity. Other considerations or goals are to reduce the comeal inflammatory response, to limit structural comeal damage, and to promote comeal reepithelialization. As is the case in other organ systems, healing of a comeal ulcer is often accompanied by neovascularization. In the eye, neovascularization and scarring are particularly deleterious as vision is dependent upon a clear cornea which requires the maintenance of the highly organized fibrin structure. Immunosuppressant corticosteroids can be used to inhibit the vessel formation but many ophthalmologists would rather not risk this indiscriminate type of immune suppression while the cornea is vulnerable due to ulceration. Thus, there exists a need in the art for agents with therapeutic efficacy in reduction of neovascularization and scanning but without the generalized immune suppressing effects of steroids. Even with current antibiotic and steroid therapies, major concerns regarding the treatment of infectious comeal ulcers remain, including: broad spectrum application; fear of antibiotic resistant strains of microbes; controversy regarding prophylactic versus therapeutic treatment of suspected infectious ulcers; non-compliant patients; control of neovascularization and scar formation. There exists a need for new therapeutic agents that would better address these issues. BPI is a protein isolated from the granules of mammalian polymorphonuclear leukocytes (PMNs or neutrophils), which are blood cells essential in the defense against invading microorganisms. Human BPI protein has been isolated from PMNs by acid extraction combined with either ion exchange chromatography [Elsbach, J. Biol. Chem. , 254: 11000 (1979)] or E. coli affinity chromatography [Weiss, et al. , Blood, 69:652 (1987)]. BPI obtained in such a manner is referred to herein as natural BPI and has been shown to have potent bactericidal activity against a broad spectrum of gram- negative bacteria. The molecular weight of human BPI is approximately 55,000 daltons (55 kD). The amino acid sequence of the entire human BPI protein and the nucleic acid sequence of DNA encoding the protein have been reported in Figure 1 of Gray et al. , J. Biol. Chem. , 264:9505 (1989), incorporated herein by reference. The Gray et al. amino acid sequence is set out in SEQ ID NO: 1 hereto. BPI is a strongly cationic protein. The N-terminal half of BPI accounts for the high net positive charge; the C-terminal half of the molecule has a net charge of -3. [Elsbach and Weiss (1981), supra. A proteolytic N- terminal fragment of BPI having a molecular weight of about 25 kD has an amphipathic character, containing alternating hydrophobic and hydrophilic regions. This N-terminal fragment of human BPI possesses the anti-bacterial efficacy of the naturally-derived 55 kD human BPI holoprotein. [Ooi et al. , J. Bio. Chem. , 262: 14891-14894 (1987)]. In contrast to the N-terminal portion, the C-terminal region of the isolated human BPI protein displays only slightly detectable anti-bacterial activity against gram-negative organisms. [Ooi et al. , J. Exp. Med. , 74:649 (1991).] An N-terminal BPI fragment of approximately 23 kD, referred to as "rBPI 23 ," has been produced by recombinant means and also retains an ti -bacterial activity against gram- negative organisms. Gazzano-Santoro et al., Infect, lmmun. 60:4754-4761 (1992). There continues to exist a need in the art for new methods and materials for treatment of co eal injury, including infection or ulceration. Products and methods responsive to this need would ideally involve substantially non-toxic, non-irritating ophthalmic preparations available in suitable amounts by means of synthetic or recombinant methods. Ideal compounds would be capable of penetrating co eal tissue and would prevent or reduce the number and severity of adverse effects, complications or conditions associated with or resulting from comeal injury. Alternatively, or in addition, such ideal compounds would enhance the effect of, or reduce the need for, other concurrently administered anti-inflammatory and/or antimicrobial therapeutic agents. SUMMARY OF THE INVENTION The present invention provides novel methods of treating co eal epithelial injury associated infection comprising topical application to the cornea of a subject having a co eal epithelial injury a bactericidal/permeability-increasing (BPI) protein product in an amount effective to reduce hyperemia, chemosis, neovascularization, mucous discharge or ulcer formation. Methods according to the invention are thus useful for reducing the adverse effects, complications or conditions associated with or resulting from a co eal injury including, comeal infection or ulceration, by topically administering a therapeutically effective amount of an ophthalmic preparation of a BPI protein product to a subject suffering from the effects of such comeal infection, ulceration or injury. The invention derives in part from the surprising discovery that topically administered BPI protein products penetrate the cornea and prevent or reduce adverse effects associated with comeal infections and ulcerations. These adverse effects include hyperemia, chemosis, mucous discharge, tearing, photophobia, keratitis, neovascularization, ulcer formation, opacification (clouding), contrast sensitivity, scarring, pain or loss of visual acuity. Confirmation of beneficial effects of practice of the invention is provided by standard ophthalmological examination including, for example, slit lamp biomicroscopy. Methods of the present invention contemplate administration of a BPI protein product in ophthalmologically acceptable preparations which may include, or be concurrently administered with, anti-inflammatory agents such as corticosteroids and/or antimicrobial agents such as ciprofloxacin gentamicin, ofloxacin and anti-fungal agents. Presently preferred BPI protein products of the invention include biologically active amino terminal fragments of the BPI holoprotein, recombinant products such as rBPI 21 and rBPI 42 and recombinant or chemically synthesized BPI-derived peptides as described in detail below. The invention further provides for the use of a BPI protein products for manufacture of a topical medicament for reducing the above- noted adverse effects, complications or conditions, associated with or resulting from co eal infection and ulceration. Numerous additional aspects and advantages of the invention will become apparent to those skilled in the art upon considering the following detailed description of the invention, which describes the presently preferred embodiments thereof, reference being made to the drawing wherein: Figure 1 is a photograph of a "control" rabbit eye 72 hours after co eal epithelium puncture and injection with Pseudomonas aeruginosa wherein post-injection treatments included an ophthalmic product vehicle solution only; and Figure 2 is a photograph of a rabbit eye 72 hours after comeal epithelium puncture and injection with Pseudomonas aeruginosa wherein the cornea was treated according to the present invention. DETAILED DESCRIPTION OF THE INVENTION Incorporated by reference herein are the disclosures of the applicant's co-owned, co-pending, concurrently-filed U.S. Patent Application Serial No. (Attorney Docket No. 27129/33007) entitled "Methods of Treating Conditions Associated With Comeal Transplantation." The present invention relates to the surprising discovery that a bactericidal/permeability-increasing (BPI) protein product can be topically administered to the cornea, in an amount effective to reduce hyperemia, chemosis, neovascularization, mucous discharge or ulcer formation associated with or resulting from comeal epithelial injury associated infection. Methods according to the invention are useful for treating subjects suffering from comeal infection, ulceration, or injury, and conditions associated therewith or resulting therefrom. Particularly valuable is the lack of comeal tissue toxicity and the effectiveness of such topically administered BPI protein products, given that penetration of comeal tissue is a necessary but not sufficient step for therapeutic efficacy. BPI protein products are shown herein to prevent or reduce adverse effects of co eal injury associated infection and ulceration including, for example, preventing or reducing hyperemia, chemosis, mucous discharge, tearing, photophobia, keratitis, neovascularization, ulcer formation (i.e. , prevent ulcer development or reduce ulcer size) opacification (clouding), contrast sensitivity, scanning, pain and loss of visual acuity as measured by standard ophthalmological examination, using, slit lamp biomicroscopy to note clinical manifestations. According to one aspect of the invention, suitable ophthalmic preparations of BPI protein product alone, in an amount sufficient for monotherapeutic effectiveness, may be administered to a subject suffering from comeal infection, ulceration, or injury, and conditions associated therewith or resulting therefrom. When used to describe administration of BPI protein product alone, the term "amount sufficient for monotherapeutic effectiveness" means a suitable ophthalmic preparation having an amount of BPI protein product that provides beneficial effects, including anti-microbial and/or anti-angiogenic effects, when administered as a monotherapy. The invention utilizes any of the large variety of BPI protein products known to the art including natural BPI protein isolates, recombinant BPI protein, BPI fragments, BPI analogs, BPI variants, and BPI-derived peptides. According to another aspect of the invention, a patient may be treated by concurrent administration of suitable ophthalmic preparations of a BPI protein product in an amount sufficient for combinative therapeutic effectiveness and one or more immunosuppressant corticosteroids in amounts sufficient for combinative therapeutic effectiveness. This aspect of the invention contemplates concurrent administration of BPI protein product with any corticosteroid or combinations of corticosteroids, including prednisolone and dexamethasone and contemplates that, where corticosteroid therapy is required, lesser amounts will be needed and/or that there will be a reduction in the duration of treatment. According to another aspect of the invention, a subject suffering from comeal epithelial injury associated infection or ulceration, and conditions associated therewith or resulting therefrom, may be treated by concurrent administration of suitable ophthalmic preparations of a BPI protein product in an amount sufficient for combinative therapeutic effectiveness and one or more antibiotics in amounts sufficient for combinative therapeutic effectiveness. This aspect of the invention contemplates concurrent administration of BPI protein product with any antimicrobial agent or combinations thereof for topical use in the eye including: antibacterial agents such as gentamicin, tobramycin, bacitracin, chloramphenicol, ciprofloxacin, ofloxacin, norfloxacin, erythromycin, bacitracin/neomycin/polymyxin B, sulfisoxazole, sulfacetamide, tetracycline, polymyxin/bacitracin, trimethroprim/polymyxin B, vancomycin, clindamycin, ticarcillin, penicillin, oxacillin or cefazolin; antifungal agents such as amphotericin B, nystatin, natamycin (pimaricin), miconazole, ketocanozole or fluconazole; antiviral agents such as idoxuridine, vidarabine or trifluridine; and antiprotozoal agents such as propamidine, neomycin, clotrimazol, miconazole, itraconazole or polyhexamethylene biguanide. This aspect of the invention is based on the improved therapeutic effectiveness of suitable ophthalmic preparations of BPI protein products with antibiotics, e.g., by increasing the antibiotic susceptibility of infecting organisms to a reduced dosage of antibiotics providing benefits in reduction of cost of antibiotic therapy and/or reduction of risk of toxic responses to antibiotics. BPI protein products may lower the minimum concentration of antibiotics needed to inhibit in vitro growth of organisms at 24 hours. In cases where BPI protein product does not affect growth at 24 hours, BPI protein product may potentiate the early bactericidal effect of antibiotics in vitro at 0-7 hours. The BPI protein products may exert these effects even on organisms that are not susceptible to the direct bactericidal or growth inhibitory effects of BPI protein product alone. This aspect of the invention is correlated to effective reversal of the antibiotic resistance of an organism by administration of a BPI protein product and antibiotic. BPI protein products may reduce the minimum inhibitory concentration of antibiotics from a level within the clinically resistant range to a level within the clinically susceptible range. BPI protein products thus may convert normally antibiotic-resistant organisms into antibiotic-susceptible organisms. According to these aspects of the invention, suitable ophthalmic preparations of the BPI protein product along with corticosteroids and/or antibiotics are concurrently administered in amounts sufficient for combinative therapeutic effectiveness. When used to describe administration of a suitable ophthalmic preparation of BPI protein product in conjunction with a corticosteroid, the term "amount sufficient for combinative therapeutic effectiveness" with respect to the BPI protein product means at least an amount effective to reduce or minimize neovascularization and the term "amount sufficient for combinative therapeutic effectiveness" with respect to a corticosteroid means at least an amount of the corticosteroid that reduces or minimizes inflammation when administered in conjunction with that amount of BPI protein product. Either the BPI protein product or the corticosteroid, or both, may be administered in an amount below the level required for monotherapeutic effectiveness against adverse effects associated with or resulting from co eal injury associated infection/ulceration. When used to describe administration of a suitable ophthalmic preparation of BPI protein product in conjunction with an antimicrobial, the term "amount sufficient for combinative therapeutic effectiveness" with respect to the BPI protein product means at least an amount effective to reduce neovascularization and/or increase the susceptibility of the organism to the antimicrobial, and the term "amount sufficient for combinative therapeutic effectiveness" with respect to an antimicrobial means at least an amount of the antimicrobial that produces bactericidal or growth inhibitory effects when administered in conjunction with that amount of BPI protein product. Either the BPI protein product or the antimicrobial, or both, may be administered in an amount below the level required for monotherapeutic effectiveness. BPI protein product may be administered in addition to standard therapy and is preferably incorporated into the care given the patient exposed to risk of comeal epithelium injury or actually suffering such injury. Treatment with BPI protein product is preferably continued for at least 1 to 30 days, and potentially longer if necessary, in dosage amounts (e.g. , dropwise administration of about 10 to about 200 μh solution of a BPI protein product at about 1 to 2 mg/mL) determined by good medical practice based on the clinical condition of the individual patient. Suitable ophthalmic preparations of BPI protein products may provide benefits as a result of their ability to neutralize heparin and their ability to inhibit heparin-dependent angiogenesis. The anti-angiogenic properties of BPI have been described in Little et al., co-owned, co-pending U.S. Application Serial No. 08/435,855 and co-owned U.S. Patent No. 5,348,942, both incorporated by reference herein. Suitable ophthalmic preparations of BPI protein products may provide additional benefits as a result of their ability to neutralize endotoxin associated with gram-negative bacteria and/or endotoxin released by antibiotic treatment of patients with comeal infection/ulceration. Suitable ophthalmic preparations of BPI protein products could provide further benefits due to their anti-bacterial activity against susceptible bacteria and fungi, and their ability to enhance the therapeutic effectiveness of antibiotics and anti-fungal agents. See, e.g., Horwitz et al., co-owned, co-pending U.S. Application Serial No. 08/372,783, filed January 13, 1995 as a continuation-in-part of U.S. Application Serial No. 08/274,299, filed July 11, 1994, which are all incorporated herein by reference and which describe BPI protein product activity in relation to gram-positive bacteria; and Little et al., co-owned, co- pending U.S. Application Serial No. 08/372,105, filed January 13, 1995 as a continuation-in-part of U.S. Application Serial No. 08/273,540, filed July 11, 1994, which are all incorporated herein by reference and which describe BPI protein product activity in relation to fungi. For ophthalmic uses as described herein, the BPI protein product is preferably administered topically, to the comeal wound or injury. Topical routes include administration preferably in the form of ophthalmic drops, ointments, gels or salves. Other topical routes include irrigation fluids (for, e.g., irrigation of wounds). Those skilled in the art can readily optimize effective ophthalmic dosages and administration regimens for the BPI protein products. As used herein, "BPI protein product" includes naturally and recombinantly produced BPI protein; natural, synthetic, and recombinant biologically active polypeptide fragments of BPI protein; biologically active polypeptide variants of BPI protein or fragments thereof, including hybrid fusion proteins and dimers; biologically active polypeptide analogs of BPI protein or fragments or variants thereof, including cysteine-substituted analogs; and BPI-derived peptides. The BPI protein products administered according to this invention may be generated and/or isolated by any means known in the art. U.S. Patent No. 5, 198,541, the disclosure of which is incorporated herein by reference, discloses recombinant genes encoding and methods for expression of BPI proteins including recombinant BPI holoprotein, referred to as rBPI 50 or ΓBPI 55 and recombinant fragments of BPI. Co-owned, copending U.S. Patent Application Ser. No. 07/885,501 and a continuation-in-part thereof, U.S. Patent Application Ser. No. 08/072,063 filed May 19, 1993 and corresponding PCT Application No. 93/04752 filed May 19, 1993, which are all incorporated herein by reference, disclose novel methods for the purification of recombinant BPI protein products expressed in and secreted from genetically transformed mammalian host cells in culture and discloses how one may produce large quantities of recombinant BPI products suitable for incorporation into stable, homogeneous pharmaceutical preparations. Biologically active fragments of BPI (BPI fragments) include biologically active molecules that have the same or similar amino acid sequence as a natural human BPI holoprotein, except that the fragment molecule lacks amino-terminal amino acids, internal amino acids, and/or carboxy-terminal amino acids of the holoprotein. Nonlimiting examples of such fragments include a N-terminal fragment of natural human BPI of approximately 25 kD, described in Ooi et al., J. Exp. Med. , 174:649 (1991), and the recombinant expression product of DNA encoding N-terminal amino acids from 1 to about 193 or 199 of natural human BPI, described in Gazzano-Santoro et al. , Infect. Immun. (50:4754-4761 (1992), and referred to as rBP^. In that publication, an expression vector was used as a source of DNA encoding a recombinant expression product (rBPI^) having the 31- residue signal sequence and the first 199 amino acids of the N-terminus of the mature human BPI, as set out in Figure 1 of Gray et al. , supra, except that valine at position 151 is specified by GTG rather than GTC and residue 185 is glutamic acid (specified by GAG) rather than lysine (specified by AAG). Recombinant holoprotein (rBPI) has also been produced having the sequence (SEQ ID NOS: 1 and 2) set out in Figure 1 of Gray et al., supra, with the exceptions noted for rBPI 23 and with the exception that residue 417 is alanine (specified by GCT) rather than valine (specified by GTT). Other examples include dimeric forms of BPI fragments, as described in co-owned and co- pending U.S. Patent No. 5,447,913 the disclosures of which are incorporated herein by reference. Preferred dimeric products include dimeric BPI protein products wherein the monomers are amino-terminal BPI fragments having the N-terminal residues from about 1 to 175 to about 1 to 199 of BPI holoprotein. A particularly preferred dimeric product is the dimeric form of the BPI fragment having N-terminal residues 1 through 193, designated rBPI 42 dimer. Biologically active variants of BPI (BPI variants) include but are not limited to recombinant hybrid fusion proteins, comprising BPI holoprotein or biologically active fragment thereof and at least a portion of at least one other polypeptide, and dimeric forms of BPI variants. Examples of such hybrid fusion proteins and dimeric forms are described by Theofan et al. in co-owned, copending U.S. Patent Application Serial No. 07/885,911, and a continuation-in-part application thereof, U.S. Patent Application Serial No. 08/064,693 filed May 19, 1993 and corresponding PCT Application No. US93/04754 filed May 19, 1993, which are all incorporated herein by reference and include hybrid fusion proteins comprising, at the amino-terminal end, a BPI protein or a biologically active fragment thereof and, at the carboxy-terminal end, at least one constant domain of an immunoglobulin heavy chain or allelic variant thereof. Biologically active analogs of BPI (BPI analogs) include but are not limited to BPI protein products wherein one or more amino acid residues have been replaced by a different amino acid. For example, co-owned, U.S. Patent No. 5,420,019 and corresponding PCT Application No. US94/01235 filed February 2, 1994, the disclosures of which are incorporated herein by reference, discloses polypeptide analogs of BPI and BPI fragments wherein a cysteine residue is replaced by a different amino acid. A preferred BPI protein product described by this application is the expression product of DNA encoding from amino acid 1 to approximately 193 (particularly preferred) or 199 of the N-terminal amino acids of BPI holoprotein, but wherein the cysteine at residue number 132 is substituted with alanine and is designated rBPI 21 Δcys or rBPI 2 ι. Other examples include dimeric forms of BPI analogs; e.g. co-owned and co-pending U.S. Patent Application Serial No. 08/212,132 filed March 11, 1994, the disclosures of which are incorporated herein by reference. Other BPI protein products useful according to the methods of the invention are peptides derived from or based on BPI produced by synthetic or recombinant means (BPI-derived peptides), such as those described in PCT Application No. US95/09262 filed July 20, 1995 corresponding to co-owned and copending U.S. Application Serial No. 08/504,841 filed July 20, 1995, PCT Application No. US94/10427 filed September 15, 1994, which corresponds to U.S. Patent Application Serial No. 08/306,473 filed September 15, 1994, and PCT Application No. US94/02465 filed March 11, 1994, which corresponds to U.S. Patent Application Serial No. 08/209,762, filed March 11, 1994, which is a continuation-in-part of U.S. Patent Application Serial No. 08/183,222, filed January 14, 1994, which is a continuation-in-part of U.S. Patent Application Ser. No. 08/093,202 filed July 15, 1993 (for which the corresponding international application is PCT Application No. US94/02401 filed March 11 , 1994), which is a continuation-in-part of U.S. Patent Application Ser. No. 08/030,644 filed March 12, 1993, the disclosures of all of which are incorporated herein by reference. The safety of BPI protein products for systemic administration to humans has been established healthy volunteers and in human experimental endotoxemia studies published in von der Mohlen et al. , Blood, 85(12):3431- 3343 (1995) and von der Mohlen et al. , J. Infect. Dis. , 772:144-151 (1995). Presently preferred BPI protein products include recombinantly- produced N-terminal fragments of BPI, especially those having a molecular weight of approximately between 21 to 25 kD such as rBPI 21 or rBPI 23 ; or dimeric forms of these N-terminal fragments (e.g., rBPI 42 dimer). Additionally, preferred BPI protein products include rBPI 55 and BPI-derived peptides. Presently most preferred is the rBPI 21 protein product. The administration of BPI protein products is preferably accomplished with a pharmaceutical composition comprising a BPI protein product and a pharmaceutically acceptable diluent, adjuvant, or carrier. The BPI protein product may be administered without or in conjunction with known surfactants, other chemotherapeutic agents or additional known antimicrobial agents. Presently preferred pharmaceutical compositions containing BPI protein products (i.e. , rBPI 21 ) comprise the BPI protein product at a concentration of 2 mg/ml in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 403 (Pluronic P123, BASF Wyandotte, Parsippany, New Jersey) (most preferred) or 0.2% poloxamer 333 (Pluronic P103 BASF Wyandotte, Parsippany, New Jersey) and 0.002% polysorbate 80 (Tween 80, ICI Americas Inc., Wilmington, Delaware). Compositions of BPI protein product and anti-bacterial activity-enhancing poloxamer surfactants are described in co- owned, co-pending U.S. Patent Application Serial Nos. 08/372, 104 filed January 13, 1995 and 08/530,599 filed September 19, 1995 the disclosures of which are incorporated herein by reference. Another pharmaceutical composition containing BPI protein products (i.e. , rBPI 21 ) comprises the BPI protein product at a concentration of 2 mg/ml in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 188 (Pluronic F 68, BASF Wyandotte, Parsippany, New Jersey) and 0.002% polysorbate 80. Yet another pharmaceutical composition containing BPI protein products (e.g., rBPI 55 , rBPI 42 , ΓBPI^) comprises the BPI protein product at a concentration of 1 mg/ml in citrate buffered saline (5 or 20 mM citrate, 150 mM NaCl, pH 5.0) comprising 0.1 % by weight of poloxamer 188 (Pluronic F-68, BASF Wyandotte, Parsippany, NJ) and 0.002% by weight of polysorbate 80 (Tween 80, ICI Americas Inc. , Wilmington, DE). Such combinations are described in co-owned, co-pending PCT Application No. US94/01239 filed February 2, 1994, which corresponds to U.S. Patent Application Ser. No. 08/190,869 filed February 2, 1994 and U.S. Patent Application Ser. No. 08/012,360 filed February 2, 1993, the disclosures of all of which are incorporated herein by reference. Other aspects and advantages of the present invention will be apparent upon consideration of the following illustrative examples wherein: Example 1 addresses the effects of various BPI protein products with respect to Pseudomonas infection in a comeal infection/ulceration rabbit model; Example 2 addresses the effects of varying formulations of a single BPI protein product with respect to Pseudomonas infection in a comeal infection/ulceration rabbit model; Example 3 addresses the effects of BPI protein product administration on Pseudomonas infection in a co eal infection/ulceration rabbit model either alone and in co-administration with various antibiotics. EXAMPLE 1 EFFECT OF BPI PROTEIN PRODUCTS ON PSEUDOMONAS INFECTION IN A CORNEAL ULCERATION RABBIT MODEL The effects of various BPI protein products were first evaluated in the context of administration both prior to and after Pseudomonas infection in a comeal infection/ulceration rabbit model. BPI protein products tested included: rBPI 42 (Expt. 1), rBPI 2I in a formulation with poloxamer 188 (Expt. 2), an anti-angiogenic BPI-derived peptide designated XMP.112 (Expt. 3), an anti-bacterial BPI-derived peptide designated XMP.105 (Expt. 4) and rBPI 2I in a formulation with poloxamer 403 (Expt. 5). The structure of XMP.112 and XMP.105 are set out in previously-noted PCT Application No. 94/02465. For these experiments, the infectious organism was a strain of Pseudomonas aeruginosa 19660 obtained from the American Type Culture Collection (ATCC, Rockville, MD). The freeze dried organism was resuspended in nutrient broth (Difco, Detroit, MI) and grown at 37 °C with shaking for 18 hours. The culture was centrifuged following the incubation in order to harvest and wash the pellet. The washed organism was Gram stained in order to confirm purity of the culture. A second generation was cultured using the same techniques as described above. Second generation cell suspensions were diluted in nutrient broth and adjusted to an absorbance of 1.524 at 600 nm, a concentration of approximately 6.55 X IO 9 CFU/ml. A final 1.3 X IO 6 fold dilution in nutrient broth yielded 5000 CFU/mL or 1.0 X 10 2 CFU/0.02 mL. Plate counts for CFU determinations were made by applying 100 μL of the diluted cell suspension to nutrient agar plates and incubating them for 24-48 hours at 37 "C. For these experiments, the animals used were New Zealand White rabbits, maintained in rigid accordance to both SERI guidelines and the ARVO Resolution on the Use of Animals in Research. A baseline examination of all eyes was conducted prior to injection in order to determine ocular health. All eyes presented with mild diffuse fluorescein staining, characteristically seen in the normal rabbit eye. The health of all eyes fell within normal limits. Rabbits weighing between 2.5 and 3.0 kg were anesthetized by intramuscular injection of 0.5-0.7 mL/kg rodent cocktail (100 mg/mL ketamine, 20 mg/mL xylazine, and 10 mg/mL acepromazine). One drop of proparacaine hydrochloride (0.5% Ophthaine, Bristol-Myers Squibb) was applied to the eye prior to injection. Twenty microliters of bacterial suspension (1 X IO 2 CFU) prepared as described above was injected into the central comeal stroma of a randomly assigned eye while the other eye remained naive. Injections, simulating perforation of the comeal epithelium, were performed using a 30-gauge 1/2-inch needle and a 100 μL syringe. For the first series of experiments, a 5-day dosing regimen of BPI protein product (test drug) was as follows: on Day 0 of the study, 40 μL of test drug or vehicle control was delivered to the test eye at 2 hours (-2) and 1 hour (-1) prior to intrastromal bacterial injection (time 0), then at each of the following 10 hours (0 through +9 hrs) post-injection for a total of 12 doses (40 L/dose); on each of Days 1-4 of the study, 40 μL of test drug or vehicle control was delivered to the test eye at each of 10 hours (given at the same time each day, e.g., 8am-5pm). For Expt. 1, 9 animals were treated, 5 with rBPL, 2 (1 mg/mL in 5 mM citrate, 150 mM NaCl, 0.1 % poloxamer 188, 0.002% polysorbate 80) and 4 with buffered vehicle (5 mM citrate, 150 mM NaCl, 0.2% poloxamer 188, 0.002% polysorbate 80). For Expt. 2, 10 animals were treated, 5 with rBPI 2 , (2 mg/mL in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 188, 0.002% polysorbate 80) and 5 with buffered vehicle. For each of Expt. 3 and Expt. 4, 5 animals were treated with XMP.112 (1 mg/mL in 150 mM NaCl) and XMP.105 (1 mg/mL in 150 mM NaCl), respectively, and 5 animals with buffered vehicle. For Expt. 5, 5 animals were treated with rBPI 21 (2 mg/mL in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 403, 0.002% polysorbate 80) and 5 animals with placebo (5 mM citrate, 150 mM NaCl, 0.2% poloxamer 403, 0.002% polysorbate 80). For these experiments, eye examinations were conducted two times each day for each 5-day study via slit lamp biomicroscopy to note clinical manifestations. Conjunctival hyperemia, chemosis and tearing, mucous discharge were graded. The grading scale for hyperemia was: 0 (none); 1 (mild); 2 (moderate); and 3 (severe). The scale for grading chemosis was: 0 (none); 1 (visible in slit lamp); 2 (moderate separation); and 3 (severe ballooning). The scale for grading mucous discharge was: 0 (none) 1 slight accumulation); 2 (thickened discharge); and 3 (discrete strands). Photophobia was recorded as present or absent. Tearing was recorded as present or absent. The co eal ulcer, when present, was assessed with respect to height (mm), width (mm), and depth (% of comeal thickness). Neovascularization was graphed with respect to the affected comeal meridians. Photodocumentation was performed daily as symptoms progressed throughout the experimental procedure. At the completion of the 5-day study period, all rabbits were sacrificed via a lethal dose of sodium pentobarbital (6 grs/mL). Comeas were harvested and fixed in half-strength Kamovsky's fixative. The comeas were processed for light microscopy using Gram stain to assay for the presence of microbial organisms and using hematoxylin and eosin to assay for cellular infiltrate. Examinations were conducted after injection of Pseudomonas at 4, 24, 28, 48, 52, 72, 76, and 96 hours for these experiments. Additional examinations were conducted at 100 and 168 hours for Expt 3 with XMP.112 since neovascularization progressed more slowly in this experiment than it did in others. The results of these examinations are reported in Table 1 for Expt. 5 wherein the BPI protein product tested (rBPI 21 , in a formulation with poloxamer 403) provided the most potent effects. TABLE 1 Mean scores of clinical observations graded on a scale of 0 (none) to 3 (severe). The results set out in Table 1 reveal that treatment of the eye prior to and after perforation injury and injection of Pseudomonas provided substantial benefits in terms of reduced hyperemia, chemosis and mucous formation, as well as reduction in incidence of neovascularization along with reduced incidence and severity of comeal ulceration. At four hours after Pseudomonas injection, fluorescein staining of the cornea in both treated and control animals revealed small areas of staining consistent with the injection (puncture) injury. At 28 hours after injection, the rBPI 21 treated eye evidenced clear ocular surfaces and typically were free of evidence of hyperemia, chemosis and mucous discharge while the vehicle treated eyes showed clouding of the ocular surface resulting from comeal edema and infiltration of white cells. Iritis was conspicuous in the vehicle treated eyes at 28 hours after injection and fluorescein dye application typically revealed areas of devitalized epithelium; severe hyperemia and moderate to severe chemosis and mucous discharge were additionally noted. At 48 hours after injection, mild hyperemia was sometimes noted in the rBPI 21 treated eyes but mucous discharge and chemosis were absent; the rBPI 2I treated comeas were otherwise typically clear and healthy appearing, as evidenced by the application of fluorescein dye. Vehicle treated eyes at 48 hours post infection displayed severe hyperemia, chemosis and mucous discharge were present; some comeas displayed comeal melting and thinning along with an ulcerating area clouded as a result of edema, cellular infiltration and fibrin deposition. At 52 hours following injection, rBPI 21 treated eyes exhibited clear and healthy comeas which resisted staining with fluorescein, indicating that the formulation is safe and non-toxic to the comeal epithelium. In vehicle treated eyes at 52 hours post infection, sloughing of comeal epithelium was evident and while chemosis was decreasing, hyperemia was severe. In these experiments, several vehicle treated eyes presented with neovascularization, with vessels growing inward toward the central cornea. This manifestation > was not noted in any rBPI 21 treated eye. Pathohistological evaluation of the rBPI 21 treated comeas stained with hematoxylin and eosin revealed healthy, intact co eal epithelium and stroma; the tissue was free of white cell infiltration. In contrast, evaluation of the vehicle treated comeas revealed absence of an epithelium and extensive infiltration of white cells into the comeal stroma. Additional pathohistological evaluation of the rBPI 21 treated comeas stained with toluidine blue also revealed healthy, intact comeal epithelium and stoma, and further revealed comeal tissue free of Pseudomonas organisms. In contrast, evaluation of the vehicle treated comeas revealed rod shaped Pseudomonas organisms in the tissue and the presence of white cells advancing toward the organisms in the tissue. These results indicate effective comeal penetration of the rBPI 21 and effective sterilization of the tissue without neovascularization. Figures 1 and 2 respectively provide a photographic comparison of representative control (placebo) and treated (rBPI 21 /poloxamer 403) results at 72 hours. The fluorescein stained treated eye (Figure 2) is healthy and clear; no keratitis is evident, confirming safety of chronic use in rabbits. In the "control" eye shown, the perithelium has severely melted; the thinning central cornea is ready to perforate. Severe hyperemia and moderate mucous discharge is apparent. Chemosis was not evident. The rBPI 21 formulation with poloxamer 403 tested in these experiments achieved the most dramatic beneficial antimicrobial and anti- angiogenic effects when compared with those of the other BPI protein product formulations tested in this severe Pseudomonas injury/infection rabbit model. Benefits in terms of suppression of neovascularization were noted for treatment with the rBPI 42 , rBPI 21 (with poloxamer 188) and XMP.112 preparations whereas treatment with XMP.105 resulted in one of the five treated eyes showing neovascularization as opposed to none for the vehicle treated animals. Further, no significant effects in reduction of hyperemia, chemosis, mucous formation and tearing were noted. The contrast in efficacy of the BPI 2 ,/poloxamer 403 results (Expt. 5) with the lesser efficacy of the other products and formulations in that study suggested that formulation components, dosage and dosage regimen for a particular BPI protein product may all have a significant role in optimizing beneficial effects associated with practice of the invention. The following Example illustrates practice of routine procedures designed to assess, in part, effects of formulation components and dosage regimens on optimization of beneficial effects attending practice of the present invention. EXAMPLE 2 EFFECT OF BPI PROTEIN PRODUCT FORMULATIONS AND DOSING ON PSEUDOMONAS INFECTION IN A CORNEAL ULCERATION RABBIT MODEL The effect of BPI protein product administration following Pseudomonas infection was evaluated in a comeal infection/ulceration rabbit model using rBPI 21 in various formulations with (A) poloxamer 188, (B) poloxamer 333, and (C) poloxamer 403 (as in Expt. 5 of Example 1). For these experiments, the infectious organism was a strain of Pseudomonas aeruginosa 19660 prepared and used to inject rabbits as described in Example 1. In a first set of studies, no beneficial effects were observed when the test product dosing regimen included no pre-injection doses of BPI protein product and treatment was withheld until commencement of ulcer formation at about 12-16 hours after the bacterial injection. Briefly put, the dosing regimen of BPI protein product employed was not sufficient to overcome the massive destructive effects of large numbers of microorganisms, where the infection was allowed to develop for 12-16 hours before intervention. In a second variant dosing and formulation study, the dosing regimen was as described in Example 1 except that animals were not dosed at 2 hours and 1 hour prior to injection with Pseudomonas, but were dosed at the time of injection and then each hour for 12 hours on the first day of the 5 day experiment. Treatment was as in Example 1 for days 2-5. For these experiments, animals were treated as follows: 5 with rBPI 21 formulated with poloxamer 188 (formulation A: 2 mg/mL rBPI 21 in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 188, 0.002% polysorbate 80), 5 with rBPI 2 , formulated with poloxamer 333 (formulation B: 2 mg/mL rBPI 2 , in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 333, 0.002 polysorbate 80), 5 with rBPI 21 formulated with poloxamer 403 (formulation C: 2 mg/mL rBPI 21 in 5 mM citrate, 150 mM NaCl, 0.2% poloxamer 403, 0.002% polysorbate 80) and 5 with phosphate buffered saline (PBS) control. Eye examinations were carried out as described in Example 1 and the animals sacrificed at the end of the 5 day protocol. Formulation C treated eyes exhibited less hyperemia than saline treated eyes up to the 28 hour evaluation. The effect was less at the 28 hour evaluation, while subsequent hyperemia scores were equivalent between test and control groups. Formulation C also consistently presented lower hyperemia scores than formulation A and B, suggesting that eyes treated with formulation C were not eliciting as much of an inflammatory response as observed the eyes in the other treated groups. Formulation C also elicited significantly lower scores for chemosis than control at the 28 hour evaluation. This effect was less at the 24 hour evaluation. Clinical scores for chemosis were consistently lower for group C than any of the other treated groups. As hyperemia increases, the vessels become progressively permeable, allowing increased serum deposition into the tissues. The formulation C treated eyes, which elicited the lowest degree of hyperemia, presented the lowest degree of chemosis. During the first 28 hours of the study, formulation C treated eyes presented consistently lower mucous discharge scores than all other groups. Neutrophil containing mucous is generally produced in response to irritation. Control treated eyes produced markedly greater mucous discharge during the first 28 hours of the study than any of the active treated groups, indicating a high degree of distress. Formulation C treated eyes displayed the smallest ulcers during the first 28 hours of the study, and in accordance with the other clinical data, was the most effective antimicrobial agent of the three formulations tested. Formulation B achieved beneficial results superior to formulation A with respect to bactericidal capability, although the differences were less than that between formulations A and C. All eyes, however, were overwhelmed by the Pseudomonas over the 28 to 48 hour period. In these experiments, formulation C demonstrated potent antimicrobial properties and was able to suppress ulcer progression. EXAMPLE 3 EFFECT OF ADMINISTRATION OF BPI PROTEIN PRODUCT AND ANTIBIOTIC FOR PSEUDOMONAS INFECTION IN A CORNEAL ULCERATION RABBIT MODEL The effect of BPI protein product administration for Pseudomonas infection is evaluated in a comeal infection/ulceration rabbit model using a BPI protein product, such as rBPI 21 , in various formulations alone and in co-administration with various antibiotics. Experiments are performed as described in Examples 1 and 2, but wherein the BPI protein product is administered as an adjunct to antibiotic treatment. Experiments are performed as described in Examples 1 and 2, except that antibiotic dosing is performed in additional to dosing with BPI protein product. For these experiments, the antibiotic dose is administered before, simultaneously with, or after each dose of BPI protein product. Numerous modifications and variations of the above-described invention are expected to occur to those of skill in the art. Accordingly, only such limitations as appear in the appended claims should be placed thereon. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Scannon, Patrick J. (ii) TITLE OF INVENTION: METHODS OF TREATING CONDITIONS ASSOCIATED WITH CORNEAL INJURY (iii) NUMBER OF SEQUENCES: 2 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Marshall, O'Toole, Gerstein, Murray & Borun (B) STREET: 6300 Seara Tower, 233 South Wacker Drive (C) CITY: Chicago (D) STATE: Illinois (E) COUNTRY: United States of America (F) ZIP: 60606-6402 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Borun, Michael F. (B) REGISTRATION NUMBER: 25,447 (C) REFERENCE/DOCKET NUMBER: 27129/33006 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 312/474-6300 (B) TELEFAX: 312/474-0448 (C) TELEX: 25-3856 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1813 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 31..1491 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 124..1491 (ix) FEATURE: (A) NAME/KEY: misc feature (D) OTHER INFORMATION: "rBPI" (xi) SEQUENCE DESCRIPTION: SEQ ID Nθ:l: CAGGCCTTGA GGTTTTGGCA GCTCTGGAGG ATG AGA GAG AAC ATG GCC AGG GGC 54 Met Arg Glu Asn Met Ala Arg Gly -31 -30 -25 CCT TGC AAC GCG CCG AGA TGG GTG TCC CTG ATG GTG CTC GTC GCC ATA 102 Pro Cys Asn Ala Pro Arg Trp Val Ser Leu Met Val Leu Val Ala lie -20 -15 -10 GGC ACC GCC GTG ACA GCG GCC GTC AAC CCT GGC GTC GTG GTC AGG ATC 150 Gly Thr Ala Val Thr Ala Ala Val Asn Pro Gly Val Val Val Arg lie -5 1 5 TCC CAG AAG GGC CTG GAC TAC GCC AGC CAG CAG GGG ACG GCC GCT CTG 198 Ser Gin Lys Gly Leu Asp Tyr Ala Ser Gin Gin Gly Thr Ala Ala Leu 10 15 20 25 CAG AAG GAG CTG AAG AGG ATC AAG ATT CCT GAC TAC TCA GAC AGC TTT 246 Gin Lys Glu Leu Lys Arg lie Lye lie Pro Asp Tyr Ser Asp Ser Phe 30 35 40 AAG ATC AAG CAT CTT GGG AAG GGG CAT TAT AGC TTC TAC AGC ATG GAC 294 Lys lie Lys His Leu Gly Lys Gly His Tyr Ser Phe Tyr Ser Met Asp 45 50 55 ATC CGT GAA TTC CAG CTT CCC AGT TCC CAG ATA AGC ATG GTG CCC AAT 342 lie Arg Glu Phe Gin Leu Pro Ser Ser Gin lie Ser Met Val Pro Aβn 60 65 70 GTG GGC CTT AAG TTC TCC ATC AGC AAC GCC AAT ATC AAG ATC AGC GGG 390 Val Gly Leu Lys Phe Ser lie Ser Asn Ala Asn lie Lys lie Ser Gly 75 80 85 AAA TGG AAG GCA CAA AAG AGA TTC TTA AAA ATG AGC GGC AAT TTT GAC 438 Lys Trp Lys Ala Gin Lys Arg Phe Leu Lys Met Ser Gly Asn Phe Asp 90 95 100 105 CTG AGC ATA GAA GGC ATG TCC ATT TCG GCT GAT CTG AAG CTG GGC AGT 486 Leu Ser lie Glu Gly Met Ser lie Ser Ala Asp Leu Lys Leu Gly Ser 110 115 120 AAC CCC ACG TCA GGC AAG CCC ACC ATC ACC TGC TCC AGC TGC AGC AGC 534 Asn Pro Thr Ser Gly Lys Pro Thr lie Thr Cys Ser Ser Cys Ser Ser 125 130 135 CAC ATC AAC AGT GTC CAC GTG CAC ATC TCA AAG AGC AAA GTC GGG TGG 582 His lie Asn Ser Val His Val His lie Ser Lys Ser Lys Val Gly Trp 140 145 150 CTG ATC CAA CTC TTC CAC AAA AAA ATT GAG TCT GCG CTT CGA AAC AAG 630 Leu lie Gin Leu Phe His Lys Lys lie Glu Ser Ala Leu Arg Asn Lys 155 160 165 ATG AAC AGC CAG GTC TGC GAG AAA GTG ACC AAT TCT GTA TCC TCC AAG 678 Met Asn Ser Gin Val Cys Glu Lys Val Thr Asn Ser Val Ser Ser Lys 170 175 180 185 CTG CAA CCT TAT TTC CAG ACT CTG CCA GTA ATG ACC AAA ATA GAT TCT 726 Leu Gin Pro Tyr Phe Gin Thr Leu Pro Val Met Thr Lys lie Asp Ser 190 195 200 GTG GCT GGA ATC AAC TAT GGT CTG GTG GCA CCT CCA GCA ACC ACG GCT 774 Val Ala Gly lie Asn Tyr Gly Leu Val Ala Pro Pro Ala Thr Thr Ala 205 210 215 GAG ACC CTG GAT GTA CAG ATG AAG GGG GAG TTT TAC AGT GAG AAC CAC 822 Glu Thr Leu Asp Val Gin Met Lys Gly Glu Phe Tyr Ser Glu Asn His 220 225 230 CAC AAT CCA CCT CCC TTT GCT CCA CCA GTG ATG GAG TTT CCC GCT GCC 870 His Asn Pro Pro Pro Phe Ala Pro Pro Val Met Glu Phe Pro Ala Ala 235 240 245 CAT GAC CGC ATG GTA TAC CTG GGC CTC TCA GAC TAC TTC TTC AAC ACA 918 His Asp Arg Met Val Tyr Leu Gly Leu Ser Asp Tyr Phe Phe Asn Thr 250 255 260 265 GCC GGG CTT GTA TAC CAA GAG GCT GGG GTC TTG AAG ATG ACC CTT AGA 966 Ala Gly Leu Val Tyr Gin Glu Ala Gly Val Leu Lys Met Thr Leu Arg 270 275 280 GAT GAC ATG ATT CCA AAG GAG TCC AAA TTT CGA CTG ACA ACC AAG TTC 1014 Asp Asp Met lie Pro Lys Glu Ser Lys Phe Arg Leu Thr Thr Lys Phe 285 290 295 TTT GGA ACC TTC CTA CCT GAG GTG GCC AAG AAG TTT CCC AAC ATG AAG 1062 Phe Gly Thr Phe Leu Pro Glu Val Ala Lys Lys Phe Pro Asn Met Lys 300 305 310 ATA CAG ATC CAT GTC TCA GCC TCC ACC CCG CCA CAC CTG TCT GTG CAG 1110 lie Gin He His Val Ser Ala Ser Thr Pro Pro His Leu Ser Val Gin 315 320 325 CCC ACC GGC CTT ACC TTC TAC CCT GCC GTG GAT GTC CAG GCC TTT GCC 1158 Pro Thr Gly Leu Thr Phe Tyr Pro Ala Val Asp Val Gin Ala Phe Ala 330 335 340 345 GTC CTC CCC AAC TCC TCC CTG GCT TCC CTC TTC CTG ATT GGC ATG CAC 1206 Val Leu Pro Asn Ser Ser Leu Ala Ser Leu Phe Leu He Gly Met His 350 355 360 ACA ACT GGT TCC ATG GAG GTC AGC GCC GAG TCC AAC AGG CTT GTT GGA 1254 Thr Thr Gly Ser Met Glu Val Ser Ala Glu Ser Asn Arg Leu Val Gly 365 370 375 GAG CTC AAG CTG GAT AGG CTG CTC CTG GAA CTG AAG CAC TCA AAT ATT 1302 Glu Leu Lys Leu Asp Arg Leu Leu Leu Glu Leu Lys His Ser Asn He 380 385 390 GGC CCC TTC CCG GTT GAA TTG CTG CAG GAT ATC ATG AAC TAC ATT GTA 1350 Gly Pro Phe Pro Val Glu Leu Leu Gin Asp He Met Asn Tyr He Val 395 400 405 CCC ATT CTT GTG CTG CCC AGG GTT AAC GAG AAA CTA CAG AAA GGC TTC 1398 Pro He Leu Val Leu Pro Arg Val Asn Glu Lys Leu Gin Lys Gly Phe 410 415 420 425 CCT CTC CCG ACG CCG GCC AGA GTC CAG CTC TAC AAC GTA GTG CTT CAG 1446 Pro Leu Pro Thr Pro Ala Arg Val Gin Leu Tyr Asn Val Val Leu Gin 430 435 440 CCT CAC CAG AAC TTC CTG CTG TTC GGT GCA GAC GTT GTC TAT AAA 1491 Pro His Gin Asn Phe Leu Leu Phe Gly Ala Asp Val Val Tyr Lys 445 450 455 TGAAGGCACC AGGGGTGCCG GGGGCTGTCA GCCGCACCTG TTCCTGATGG GCTGTGGGGC 1551 ACCGGCTGCC TTTCCCCAGG GAATCCTCTC CAGATCTTAA CCAAGAGCCC CTTGCAAACT 1611 TCTTCGACTC AGATTCAGAA ATGATCTAAA CACGAGGAAA CATTATTCAT TGGAAAAGTG 1671 CATGGTGTGT ATTTTAGGGA TTATGAGCTT CTTTCAAGGG CTAAGGCTGC AGAGATATTT 1731 CCTCCAGGAA TCGTGTTTCA ATTGTAACCA AGAAATTTCC ATTTGTGCTT CATGAAAAAA 1791 AACTTCTGGT TTTTTTCATG TG 1813 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 487 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Arg Glu Asn Met Ala Arg Gly Pro Cys Asn Ala Pro Arg Trp Val -31 -30 -25 -20 Ser Leu Met Val Leu Val Ala He Gly Thr Ala Val Thr Ala Ala Val -15 -10 -5 1 Asn Pro Gly Val Val Val Arg He Ser Gin Lys Gly Leu Asp Tyr Ala 5 10 15 Ser Gin Gin Gly Thr Ala Ala Leu Gin Lys Glu Leu Lys Arg He Lys 20 25 30 He Pro Asp Tyr Ser Asp Ser Phe Lys He Lys His Leu Gly Lys Gly 35 40 45 His Tyr Ser Phe Tyr Ser Met Asp He Arg Glu Phe Gin Leu Pro Ser 50 55 60 65 Ser Gin He Ser Met Val Pro Asn Val Gly Leu Lys Phe Ser He Ser 70 75 80 Asn Ala Asn He Lys He Ser Gly Lys Trp Lys Ala Gin Lys Arg Phe 85 90 95 Leu Lys Met Ser Gly Asn Phe Asp Leu Ser He Glu Gly Met Ser He 100 105 110 Ser Ala Asp Leu Lys Leu Gly Ser Asn Pro Thr Ser Gly Lye Pro Thr 115 120 125 He Thr Cys Ser Ser Cys Ser Ser His He Asn Ser Val His Val His 130 135 140 145 He Ser Lys Ser Lys Val Gly Trp Leu He Gin Leu Phe His Lys Lys 150 155 160 Ile Glu Ser Ala Leu Arg Asn Lys Met Asn Ser Gin Val Cys Glu Lys 165 170 175 Val Thr Asn Ser Val Ser Ser Lys Leu Gin Pro Tyr Phe Gin Thr Leu 180 185 190 Pro Val Met Thr Lys He Asp Ser Val Ala Gly He Asn Tyr Gly Leu 195 200 205 Val Ala Pro Pro Ala Thr Thr Ala Glu Thr Leu Asp Val Gin Met Lys 210 215 220 225 Gly Glu Phe Tyr Ser Glu Asn His His Asn Pro Pro Pro Phe Ala Pro 230 235 240 Pro Val Met Glu Phe Pro Ala Ala His Asp Arg Met Val Tyr Leu Gly 245 250 255 Leu Ser Asp Tyr Phe Phe Asn Thr Ala Gly Leu Val Tyr Gin Glu Ala 260 265 270 Gly Val Leu Lys Met Thr Leu Arg Asp Asp Met He Pro Lys Glu Ser 275 280 285 Lys Phe Arg Leu Thr Thr Lye Phe Phe Gly Thr Phe Leu Pro Glu Val 290 295 300 305 Ala Lys Lys Phe Pro Asn Met Lys He Gin He His Val Ser Ala Ser 310 315 320 Thr Pro Pro Hie Leu Ser Val Gin Pro Thr Gly Leu Thr Phe Tyr Pro 325 330 335 Ala Val Asp Val Gin Ala Phe Ala Val Leu Pro Asn Ser Ser Leu Ala 340 345 350 Ser Leu Phe Leu He Gly Met His Thr Thr Gly Ser Met Glu Val Ser 355 360 365 Ala Glu Ser Asn Arg Leu Val Gly Glu Leu Lys Leu Asp Arg Leu Leu 370 375 380 385 Leu Glu Leu Lys His Ser Asn He Gly Pro Phe Pro Val Glu Leu Leu 390 395 400 Gin Asp He Met Aβn Tyr He Val Pro He Leu Val Leu Pro Arg Val 405 410 415 Aβn Glu Lys Leu Gin Lys Gly Phe Pro Leu Pro Thr Pro Ala Arg Val 420 425 430 Gin Leu Tyr Asn Val Val Leu Gin Pro Hie Gin Asn Phe Leu Leu Phe 435 440 445 Gly Ala Asp Val Val Tyr Lye 450 455 ***PEPTIDES???***

Claims: WHAT IS CLAIMED IS: 1. A method for treating comeal epithelium injury associated infection comprising administering to the comea of a subject having a co eal epithelium injury a bactericidal/permeability-increasing (BPI) protein product in an amount effective to reduce hyperemia, chemosis, mucous discharge, neovascularization or ulcer formation. 2. The method of claim 1 wherein the BPI protein product is an amino-terminal fragment of BPI protein. 3. The method of claim 1 wherein the BPI protein product is rBPI, The method of claim 1 wherein the BPI protein product is rBPI, The method of claim 1 wherein the BPI protein product is rBPI 42 . 6. The method of claim 1 further comprising the step of rdministering an antibiotic or anti-fungal agent. 7. The method of claim 1 further comprising the step of administering an anti-inflammatory agent.

Inventors: SCANNON PATRICK J, SCANNON, PATRICK, J.
Applicants: XOMA CORP, XOMA CORPORATION
Application year: 1996
Main classification: A61K38
IPC codes: A61K31, A61K38, A61K45, A61P27, C07K14