From: Billi Goldberg <bigoldberg@igc.apc.org>
Subject: Revised DNCB Treatment Instructions
Note: Copyright 1992, Dan R. Greening. Non-commercial reproduction allowed.
Archive-Number: 6678
Organization: unspecified
Date: Sat, 26 Dec 92 09:16:00 PST

I am reposting the instructions because of recent experiences with
DNCB users. It appears to be critical to move the application sites
to all parts of the body: arms, legs, thighs, trunk, etc. so as to
assure that DNCB T4 memory cells have infiltrated all parts of the
body. Since these memory cells are systemically activated when DNCB
is applied, this will result in the greatest intensity of non-specific
as well as specific immune responses to the DNCB immunogen. Also, 
reactions should be strong at the application site but not unduly
uncomfortable. The stronger the DNCB, the greater the number of DNCB
haptens that initate Th1 or cell mediated immune responses to DNCB.

                DNCB TREATMENT INSTRUCTIONS (Rev. 12/92) 
                       by Billi Goldberg

DESCRIPTION 

DNCB (dinitrochlorobenzene) is a simple organic compound easily made by 
chemists. It is an unpatentable and inexpensive substance that is 
usually obtained in either bulk crystal form or liquid form (in an 
acetone base) from chemical supply houses. In industry, DNCB is used in 
the processing of color photography. It is also in use as an algicide in 
air conditioners and refrigeration equipment.

In past decades, researchers discovered dermatological uses for DNCB 
that involve direct application to the skin. In the process of exploring 
the dermatological applications of DNCB, researchers also discovered 
that DNCB has a very low toxic risk.

DOSAGE AND ADMINISTRATION 

For dermatological purposes, DNCB is currently available in four 
solution strengths, namely 10%, 2%, 0.2% and 0.02%. Each solution 
has an acetone base.

The stronger 10% solution is used initially in very small amounts to 
sensitize the body to DNCB. The weaker 2%, 0.2% and 0.02% solutions, 
which are applied using a cotton swab and accommodate differences in 
sensitivity, are intended for ongoing applications.

ACTION 

The mechanism of DNCB is similar to that of poison oak. Both produce a 
redness or itching of the skin indicative of contact dermatitis, as well 
as an underlying bodily effect.

ADVERSE REACTIONS 

Depending on the solution strength, DNCB may cause either a strong 
or mild form of contact dermatitis at the application site. The 
stronger, sensitizing 10% solution may cause a more noticeable 
itchiness, tingling, burning or redness within hours of application. The 
weaker 2%, 0.2% and 0.02% solutions cause a much milder skin reaction 
similar to the redness experienced from sunburn.

WARNINGS AND PRECAUTIONS 

1. Use only the 10% solution for sensitization unless a dermatologist 
    knowledgeable in contact sensitization recommends differently.

2. If severe contact dermatitis occurs, apply itch-reducing medicines 
    (e.g. calamine lotion) or Bactine directly to the rash. The use of 
    cortisone or hydrocortisone creams is not recommended for the 
    itching.                                                        

3. If severe blistering occurs at an application site during 
    sensitization, stop further applications and treat the blistering 
    like any other minor abrasion. Use Bactine or another topical 
    antibacterial medication. Once the skin heals, resume applications 
    using a lower concentration.                                    

4. Avoid sunbathing or sustained exposure to the sun while using DNCB. 
    The absorption of the sun's rays by the skin may inhibit the 
    intended reaction.                                              

5. Do not apply DNCB directly to any kind of topical skin disease or 
    disorder, such as Kaposi Sarcoma (KS) lesions.

6. If DNCB has been previously used or the skin exposed to DNCB (for 
    example, while developing color photography), do not use the 10% 
    solution for initial sensitization. Use the weaker 2% solution or 
    consult a dermatologist.

7. To avoid spreading the applied DNCB to other parts of the body, 
    completely cover the treated area with a bandage. When the bandage 
    is removed, immediately wash the treated area with soap and water.

8. To maintain the potency of the DNCB solution, keep the solution 
    capped with an air-tight cap, refrigerated, and away from sunlight.

DIRECTIONS FOR USE

Using DNCB involves three distinct steps: Sensitization, Verification 
and Weekly treatment. Before beginning ongoing weekly treatments 
with DNCB, the body must first be sensitized to DNCB using the 10% 
solution. Then, four weeks later, verify sensitization by applying the 
2% solution and checking for a specific bodily reaction (known as a 
reflare).

If sensitization is verified, start regular treatments the following 
week. If sensitization cannot be verified after one attempt, which is 
the case for some individuals, repeat sensitization with the 10% 
solution in increments of seven days until it occurs.

The information that follows describes each of the three steps and 
explains how to verify sensitization. These are very important and 
must be followed.

STEP I - SENSITIZATION 

Complete this step with the 10% solution before starting weekly 
treatments. If sensitization is effective, perform the following only 
once.

If you have previously used DNCB or have been exposed to DNCB start only 
with the 2% solution.

1. Saturate a cotton swab (Q tip) in the 10% DNCB solution and paint 
    an area approximately 2 inches in diameter. The underside of the 
    forearms and the inside of the upper arms are best.

2. When dry, saturate the cotton swab in the 10% DNCB solution and 
    repeat the previous operation.

3. Blow on the liquid to make the DNCB evaporate more quickly 

4. Cover the application site with an adhesive bandage (Band-Aid).

5. After 8-12 hours or overnight, remove the bandage and wash the 
    application site with soap and water.

STEP II - VERIFICATION 

The presence or absence of contact dermatitis is not a valid indicator 
that the body has been sensitized to DNCB. Therefore verify the 
underlying bodily sensitization as follows by applying the 2% DNCB 
solution four weeks  after applying the 10% DNCB.

1. Saturate a cotton swab (Q tip) in the 2% DNCB solution.

2. Paint a skin area approximately two inches in diameter with the swab 
    on the arm opposite of the one used for the sensitization 
    application. The underside of the forearms and the inside of the 
    upper arms are best.

3. When the application site appears dry, paint the site again.

4. Cover the skin area with an adhesive bandage (a Band-Aid).

5. After 8-12 hours or overnight, remove the bandage and wash the 
    application site with soap and water.

6. Inspect the application site within 24 to 72 hours after applying 2% 
    DNCB solution. A typical reaction is a slight to moderate reddening, 
    itching or burning.

7. Inspect the original sensitization site where the 10% DNCB solution 
    was applied. The desired reaction--and proof of sensitization--is 
    any reddening of the original site. This reddening is a "reflare." 
    (The intensity of the reflare will be significantly less than the 
    response first achieved with the 10% DNCB solution.)

If sensitization is not achieved (that is, the original sensitization 
site does not reflare), repeat the sensitization process using the 
10% solution one week after applying the 2% solution. Keep repeating the 
sensitization process in increments of seven days with the 10% solution 
until reflare occurs.

Once reflare occurs, begin weekly treatments. It is not necessary to 
repeat verification with the 2% solution.

STEP III - WEEKLY TREATMENT 

One week after verifying sensitization in Step II, start ongoing 
treatments with the correct solution strength.

The correct solution strength is the weakest solution that causes 
meaningful reddening, itching or burning sensations at an application 
site. Try the 2%, 0.2% and 0.02% solutions to determine if one will 
produce such a skin reaction. Using a stronger solution than is required 
does not increase the effectiveness of the DNCB. On the contrary, 
ongoing use of an overly strong solution may increase discomfort and 
overstimulate the body. But, the use of too weak a solution will not 
result in the required immune response.

If the 2% solution does not result in reddening, itching or burning 
sensations at the application site, use the 10% solution weekly until 
the response is greater than appropriate. At that time start using the 
next weaker solution that results in an appropriate response. This may 
be necessary for individuals that are severely immunodepressed.

Apply weekly treatments as follows:

1. Saturate a cotton swab in the appropriate DNCB solution.

2. Paint a skin area approximately two inches in diameter with the swab.

3. When the application site appears dry, paint the site again.

4. Cover the skin area with an adhesive bandage.

5. After 8-12 hours or overnight, remove the bandage and wash the 
    application site with soap and water.

6. Inspect the application site within 24 to 72 hours after applying the 
    DNCB solution. A typical reaction is a slight to moderate reddening, 
    itching or burning at the application site. This reaction is 
    evidence of the intended bodily response.

Repeat the application in increments of seven days  applying the DNCB to 
a different skin site each time. The underside of the forearms and the 
inside of the upper arms are best, but it should be applied to the trunk 
and legs. When the application site is changed, it might take two to 
three applications at the different areas before reactions occur. This 
is due to the lack of circulating T4 DNCB memory cells at the new 
application site. When changing application sites from the arms to the 
legs or trunks, it is advisable to increase the solution strength for 
the initial application. 

After four to six weeks, or when normal skin color returns, reapply the 
DNCB to a previous application site.

If DNCB is not applied weekly, an unusually strong reaction might take 
place, even with the weakest solution, during subsequent applications. 
This is caused by suppressor T cell depletion. Just continue using the 
.02% solution on a weekly basis, and the application site dermatitis 
will lessen as time progresses. DNCB must be used on a weekly basis to 
achieve the best results.

RECOMMENDATION

A collaborative relationship with your doctor or dermatologist is highly 
recommended when using DNCB. The Treatment Instructions must be followed 
exactly unless a dermatologist experienced in contact sensitivity 
recommends differently.
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