Lysozyme (chicken egg white)

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

The skin sensitization potential of enzymes has recently been reviewed by Basketter et al. ^[6] and HERA ^[4;5] revealing that enzymes should not be considered skin sensitizers. In addition, there is an unequivocal statement from AMFEP (www.amfep.org) on this topic showing that enzymes do not have skin sensitizing potential. The lack of skin sensitizing potential is substantiated by evidence from robust human experimental data and extensive in-use human studies performed with detergents containing enzymes ^{[4 -8]}. All of these studies confirmed that the presence of enzymes in the detergents did not result in contact skin sensitization, including those conducted with atopic individuals.

However, in spite of clear evidence that enzymes should not be considered skin sensitizers, animal skin sensitization models might give rise to positive results. This is because, just like the previously used guinea pig skin sensitization models, the Local Lymph Node Assay (LLNA), (OECD Test Guideline 429) is inappropriate for the assessment of proteins. These animal models are validated for the testing of small chemicals, not for water soluble protein-based materials, known to be human respiratory allergens. The LLNA does not discriminate between chemical and respiratory sensitizers ^[9], leading to the real risk of false-positive results with proteins, particularly those already known to be sensitizing by the respiratory route, such as enzymes. Indeed, in our experience, all foreign proteins can be made to generate skin reactions in suitably treated animals, including the OECD recognized guinea pig tests and the LLNA ^[10]. This makes the available animal models inappropriate when used with proteins. Therefore, the assessment of enzymes in any of the existing animal models can be predicted not to provide new and useful knowledge. This conclusion is based on the following considerations:

- The results of predictive testing in man demonstrate that enzymes do not have skin sensitization potential for man.

- In clinical settings, enzymes have only very rarely been suggested as a possible cause of allergic contact dermatitis (ACD). Even in these few cases, a causal relationship has never been proven. Further, several clinical studies have demonstrated that enzymes are not a cause of ACD ^{[5; 8; 11 -15]}.

- ACD has never been reported in the detergent enzyme industries where there has been extensive occupational enzyme exposure which, in the past, led to respiratory sensitization and/or irritant dermatitis. For more than 40 years, billions of consumers have had regular, often daily, skin exposure to enzymes during laundry by hand but there is no evidence that this exposure has given rise to skin sensitization.

- The available skin sensitization test methods are not suitable for enzymes. No animal model has been developed or validated for assessing proteins as contact skin sensitizers. So far, no in vitro models exist either.

Since enzyme products are well documented not to be skin sensitizers in man and because no suitable animal model or in vitro assay for protein skin sensitization exists, we consider testing enzymes in animal models developed for chemical contact allergens as both scientifically and ethically unjustified. Finally, the precautions recommended in the material safety data sheets should be sufficient to prevent even a theoretical hazard of skin sensitization.

Reference

[1] Basketter,D.A., English,J.S., Wakelin,S.H., and White,I.R. (2008) Enzymes, detergents and skin: facts and fantasies. British journal of dermatology 158, 1177-1181

[2] HERA Human and environmental risk assessment on ingredients of household cleaning products - alpha-amylases, cellulases and lipases. 2005.

[3] HERA Human and environmental risk assessment on ingredients of household cleaning products - Subtilisins (Proteases). Edition 2.0. 2007.

[4] Bannan, E.A., Griffith, J.F., Nusair, T.L., and L.J. Sauers (1983) Skin testing of laundered fabrics in the dermal safety assessment of enzyme containing detergents. Journal of Toxicology - Cutaneous and Ocular Toxicology 11, 327-339

[5] Griffith, J.F., Weaver, J.E., Whitehouse, H.S., Poole, R.L., and Newmann EA Nixon, G.A. (1969) SAFETY EVALUATION OF ENZYME DETERGENTS ORAL AND CUTANEOUS TOXICITY IRRITANCY AND SKIN SENSITIZATION STUDIES. Food and Cosmetics Toxicology 7, 581-593

[6] Rodriguez,C., Calvin,G., Lally,C., and LaChapelle,J.M. (1994) Skin effects associated with wearing fabrics washed with commercial laundry detergents. Journal of Toxicology - Cutaneous and Ocular Toxicology 13, 39-45

[7] Cormier,E.M., Sarlo,K., Scott,L.A., MacKenzie,D.P., Payne,N.S., Carr,G.J., Smith,L.A., Cua-Lim,F., Bunag,F.C., and Vasunia,K. (2004) Lack of type 1 sensitization to laundry detergent enzymes among consumers in the Philippines: results of a 2-year study in atopic subjects. Annals of Allergy Asthma and Immunology 92, 549-557

[8] White,I.R., Lewis,J., and el,A.A. (1985) Possible adverse reactions to an enzyme-containing washing powder. Contact Dermatitis 13, 175-179

[9] Kimber,I., Agius,R., Basketter,D.A., Corsini,E., Cullinan,P., Dearman,R.J., Gimenez-Arnau,E., Greenwell,L., Hartung,T., Kuper,F., Maestrelli,P., Roggen,E., and Rovida,C. (2007) Chemical respiratory allergy: opportunities for hazard identification and characterisation. The report and recommendations of ECVAM workshop 60. Altern Lab Anim 35, 243-265

[10] Festersen,U., Rasmussen,C., Kjaer,T.M.R., Soni,N.K., Roggen,E.L., and Berg,N.W. (2008) Alternative application route in the LLNA provides crucial environmental enrichement and broadens the usability of vehicles. AATEX 14, 433-436

[11] Belsito,D.V., Fransway,A.F., Fowler,J.F., Jr., Sherertz,E.F., Maibach,H.I., Mark,J.G., Jr., Mathias,C.G., Rietschel,R.L., Storrs,F.J., and Nethercott,J.R. (2002) Allergic contact dermatitis to detergents: a multicenter study to assess prevalence. Journal of the American Academy of Dermatology 46, 200-206

[12] Lee,M.Y., Park,K.S., Hayashi,C., Lim,H.H., Lee,K.H., Kwak,I., and Laurie,R.D. (2002) Effects of repeated short-term skin contact with proteolytic enzymes. Contact Dermatitis 46, 75-80

[13] Pepys,J., Wells,I.D., D'souza,M.F., and Greenberg,M. (1973) CLINICAL AND IMMUNOLOGICAL RESPONSES TO ENZYMES OF BACILLUS-SUBTILIS IN FACTORY WORKERS AND CONSUMERS. Clinical Allergy 3, 143-160

[14] Peters,G., Johnson,G.Q., and Golembiewski,A. (2001) Safe use of detergent enzymes in the workplace. Appl.Occup Environ.Hyg. 16, 389-396

[15] Zachariae,H., Thomsen,K., and Rasmussen,O.G. (1973) Occupational enzyme dermatitis. Results of patch testing with Alcalase. Acta dermato-venereologica 53, 145-148

Migrated from Short description of key information:
Not expected to cause skin sensitization.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

In the EFSA review (2010 ^[1]) on the potential of microorganisms, microbial products and enzymes to induce respiratory sensitization was reported that enzymes have been a cause of concern for several decades in food and feed industry, albeit at lower levels than in the bakery or detergent industry ^[2].

The most reported cause of respiratory sensitization by enzymes in the food and feed industry is the case of alpha-amylase and baker’s asthma, but other enzymes such as proteases, cellulase and glucose oxydase are also a cause of concern. To compound the problem, industrial enzyme grades are often poor and contain other enzymes from the same production organism, leading to unexpected sensitization and cross-reactions ^[3].

In the remarks of the EFSA document ^[1] was reported that the inherent properties of an enzyme may not be decisive, as to whether it is going to create inhalation allergies and that it is not known whether sensitization always lead to clinical allergy, but based on industrial hygiene experiences and knowledge from other areas of allergy, it must be assumed that sensitization would be a very strong risk factor for also developing symptoms upon continued exposure.

In the specific case of Lysozyme some literature data of cases reported are available:

- Egg lysozyme. A case report: a worker in a company manufacturing egg lysozyme powder for use in the pharmaceutical industry showed respiratory sensitization (asthma symptoms). ^{[4 and 5]}

- Serratial peptidase and lysozyme. A case report from pharmaceutical industry: One worker sensitized(asthma symptoms). ^{[4 and 6]}

- The patient sifted 50 mg of powdered enzyme back and forth between porous trays; lactose powder as placebo test; FEV1.0 and FVC recorded; for possible late reaction, a mini-Wright flowmeter for home measurements. Symptoms: Immediate asthmatic reaction. ^{[4 and 5]}

- Interesting case reports on amylase include a double sensitization to lysozyme and amylase in a baker with rhinoconjunctivitis and asthma. ^{[1 and 8]}

Considering the high concern related to the sensitization reaction, the information available about reported cases, it is considered appropriate to evaluate Lysozyme as potentially able to cause allergy or asthma symptoms or breathing difficulties if inhaled.

Reference

[1] SCIENTIFIC / TECHNICAL REPORT submitted to EFSA. Cyril Martela, Gunnar D. Nielsenb, Adriano Maric, Tine Rask Lichta, Lars K. Poulsena. Bibliographic review on the potential of microorganisms, microbial products and enzymes to induce respiratory sensitization. CFP/EFSA/FEEDAP/2009/02. Accepted for Publication on 22nd October 2010

[2] Vanhanen M, Tuomi T, Tiikkainen U, Tupasela O, Tuomainen A, et al. (2001) Sensitisation to enzymes in the animal feed industry. Occupational and Environmental Medicine 58: 119-123.

[3] Vanhanen M, Tuomi T, Hokkanen H, Tupasela O, Tuomainen A, et al. (1996) Enzyme exposure and enzyme sensitisation in the baking industry. OccupEnvironMed 53: 670-676.

[4] Vanhanen Markku. Exposure, sensitization and allergy to industrial enzymes. People and Work Research Reports 46. Finnish Institute of Occupational Health Department of Pulmology, Helsinki University Central Hospital Helsinki 2001.

[5] Bernstein JA, Kraut A, Bernstein DI, Warrington R, Bolin T, Warren CPW, Bernstein IL. Occupational asthma induced by inhaled egg lysozyme. Chest 1993;103:532–535

[6] Park HS, Nahm DH. New occupational allergen in a pharmaceutical industry: serratial peptidase and lysozyme chloride. Ann Allergy Asthma Immunol 1997;78:225–229.

[7] Santaolalla M, De BM, De FC, Gandolfo M, Zubeldia J, et al. (2002) Double sensitization to enzymes in a baker. Allergy 57: 957

Migrated from Short description of key information:
May cause allergy or asthma symptoms or breathing difficulties if inhaled.

Justification for classification or non-classification

According to CLP regulation (EC1272/2008) Lysozyme is classified as Resp. Sens. 1, H334.