Endoplasmic Reticulum-Associated Degradation-Dependent Processing in Cross-Presentation and Its Potential for Dendritic Cell Vaccinations: A Review
Round 1
Reviewer 1 Report
General comments:
Because of translation from Japanese to English, some of the intent of content is not clear, especially in the introductory background material.
Please make it clear in text which conclusions are based only on mouse data and not verified in humans or known to be different in humans.
Should make it clear in that background section that in the absence of cross presentation, theoretically will get predominantly Th2 responses rather Th1 responses associated with antigen-specific CTL.
It would be useful for the authors to specifically address whether ex vivo approaches to mature DC are beneficial or detrimental in terms of cross presentation and trafficking to lymph nodes.
It would be useful for the authors to comment on the pros and cons of electroporation in terms of direct MHC I presentation and cross presentation.
Specific comments:
Line 30-31 suggest saying: to date therapeutic cancer vaccines have resulted in limited clinical benefit.
Line 33-34 Not correct as written. The immune system is able to activate CTLs, but they are deactivated by checkpoint inhibitors.
Line 37. DC are not usually isolated from patients. Rather monocytes are isolated and differentiated into DC.
Line 38. DC have been given by a variety of routes, not just infusions, e.g. subcutaneous, intradermal, intralesional, etc.
Line 41 suggest “via” rather than “upon” MHCII ….
Line 46. Suggest insert “ immune’ to read “effective immune response”
Line 47-48 should be revised. Suggest “However, the results of DC vaccination have been disappointing, and limited CP activity may have resulted in insufficient numbers of CTLs [1].”
Line 55-57 unclear what is intended in this sentence. What is meant by “these investigations”?—clinical trials with DC vaccines? What is meant by “partial improvement of CP efficiency?
Line 82: After detecting immunological stimulants and processing antigen, DCs undergo maturation and migrate into the draining lymph nodes …
Line 137-138. Unclear what was intended in this sentence.
Lines 197-199 abbreviate MTOR after first used in the sequence of sentences.
Line 451-452: believe authors did not intend for “not” in this sentence.
Line 464-467. Unclear what is intended here. Not sure how the role of the adjuvants mentioned “indicates” importance of DC activation and maturation. What about other adjuvants that have been used? What about coadministration with GM-CSF, locally or systemically?
484-486 unclear how “innate immunity” plays a critical role. Secretion of cytokines? Cell-cell interactions? Both?
Author Response
Response to reviewers’ comments
Reviewer 1
Because of translation from Japanese to English, some of the intent of content is not clear, especially in the introductory background material. Please make it clear in text which conclusions are based only on mouse data and not verified in humans or known to be different in humans.
→ Most of the cited studies involved experiments in mice. Although the molecular mechanisms of DCs are essentially almost equivalent between humans and mice, we have indicated the studied species when a significant difference was observed between species or when the results were shown for only one species.
Should make it clear in that background section that in the absence of cross presentation, theoretically will get predominantly Th2 responses rather Th1 responses associated with antigen-specific CTL.
→ We have clarified this by added the following sentence to the Introduction:
“On the other hand, in the absence of CP, the immune system theoretically produces predominantly T helper 2 (Th2) responses rather than T helper 1 (Th1) responses associated with antigen-specific CTLs, resulting in no tolerance to cancer.”
It would be useful for the authors to specifically address whether ex vivo approaches to mature DC are beneficial or detrimental in terms of cross presentation and trafficking to lymph nodes.
→ We have revised section 6 (DC Maturation) to address the benefit of DC maturation upon CP and trafficking to lymph nodes.
It would be useful for the authors to comment on the pros and cons of electroporation in terms of direct MHC I presentation and cross presentation.
→ We have added a paragraph in section 2.2 (DC Vaccination) describing the merits and demerits of each antigen-loading method.
Specific comments:
Line 30-31 suggest saying: to date therapeutic cancer vaccines have resulted in limited clinical benefit.
→ We have incorporated this statement as suggested.
Line 33-34 Not correct as written. The immune system is able to activate CTLs,but they are deactivated by checkpoint inhibitors.
→ Additionally, although cancer vaccines can activate cancer-specific CTL, malignant cancer cells are equipped with several methods to evade the immune system
Line 37. DC ar e not usually isolated from patients. Rather monocytes are isolated and differentiated into DC.
→ We have added a paragraph in section 2.2 (DC Vaccination) describing the isolation methods of DCs from patients.
Line 38. DC have been given by a variety of routes, not just infusions, e.g. subcutaneous, intradermal, intralesional, etc.
→ We have mentioned the various administration routes in section 6 (DC Maturation).
Line 41 suggest “via” rather than “upon” MHCII ….
→ We have corrected "upon" to "via."
Line 46. Suggest insert “immune’ to read “effective immune response”
→ We have revised “effective response” to “effective immune response.”
Line 47-48 should be revised. Suggest “However, the results of DC vaccination have been disappointing, and limited CP activity may have resulted in insufficient numbers of CTLs [1].”
→ We have revised this sentence as recommended.
Line 55-57 unclear what is intended in this sentence. What is meant by “these investigations”? —clinical trials with DC vaccines? What is meant by “partial improvement of CP efficiency?
→ We have clarified our meaning by revising this sentence as follows:
“Although one of the aims of these investigations was the improvement of CP efficiency, which was partially accomplished in a mouse model [17-19], this has not contributed to the improvement of DC vaccination in clinical trials [20].”
Line 82: After detecting immunological stimulants and processing antigen, DCs undergo maturation and migrate into the draining lymph nodes …
→ We have revised this sentence as recommended by the reviewer.
Line 137-138. Unclear what was intended in this sentence.
→ We have added section 2.2 (DC Vaccination) to more clearly explain our view.
Lines 197-199 abbreviate MTOR after first used in the sequence of sentences.
→ We have abbreviated mTOR as advised.
Line 451-452: believe authors did not intend for “not” in this sentence.
→ We have revised this sentence as follows to clarify our meaning:
“However, excessive ER stress results in inflammation or an immunosuppressive phenotype, which promotes cancer escape from the immune system. Further investigation is necessary to apply ERAD-dependent processing to antigen-loading methods.”
Line 464-467. Unclear what is intended here. Not sure how the role of the adjuvants mentioned “indicates” importance of DC activation and maturation. What about other adjuvants that have been used? What about coadministration with GM-CSF, locally or systemically?
→ In this paragraph, we did not want to discuss adjuvants in depth but rather focus on restricted types of immune modifiers that activate the innate immunity of DCs to demonstrate the importance of the innate immunity in DC vaccination. We have revised this text as shown below to clarify our meaning:
“Some immune response modifiers, such as imiquimod [181], BCG, or picibanil [182], which are approved as anti-cancer drugs, activate innate immunity and facilitate DC activation and maturation. These results indicate that the activation of innate immunity plays an essential role in activation immunotherapy, including DC vaccination.”
484-486 unclear how “innate immunity” plays a critical role. Secretion of cytokines? Cell-cell interactions? Both?
→ We have revised this sentence as follows to clarify our meaning:
“These results suggest that activation of innate immunity may cause delayed maturation of endocytic compartments with non-self-proteins [118-122], resulting in improvement of CP efficiencies of non-self-proteins in addition to the induction of DC maturation.”
Reviewer 2 Report
The authors summarized the progress of endoplasmic reticulum-associated degradation-dependent processing in cross-presentation and its potential for dendritic cell vaccinations. This manuscript provides useful information but the following comments need to be addressed.
The authors should provide a table to list the markers of DC subsets. The authors should add more information to address the role of different pathways for antigen uptake in cross-presentation. Figure 1, antigen presentation of MHC II should be added. PPRs should be changed to PRRs. Lines 197-200, the description is confused. Figure 2, ‘incorrect disulfide bods’ should be changed to ‘incorrect disulfide bonds’. There are many spelling errors.
Line 80, ‘PPRs’ should be changed to ‘PRRs’. Line 103, [31,34] should move to the end of this sentence. Line 130, -glucans should be corrected. Line 202, + should be superscript. Lines 260 and 292, M should be corrected. Line 356, ‘SEC61 and ‘ should be corrected. Line 442, ‘DCs’ should be changed to ‘DCs.’ Line 460, ‘pattern recognition receptors (PRRs)’ should be changed to PRRs. Line 464, PPRs should be change to PRRs.
Author Response
Response to reviewers’ comments
Reviewer 2
The authors summarized the progress of endoplasmic reticulum-associated degradation-dependent processing in cross-presentation and its potential for dendritic cell vaccinations. This manuscript provides useful information but the following comments need to be addressed.
The authors should provide a table to list the markers and cytokines of each the DC subset.
→ We have added Table 1, which contains a list of markers and cytokines of each DC subset, according to the reviewer's comment.
The authors should add more information to address the role of different pathways for antigen uptake in cross-presentation.
→ We have added a paragraph in section 2.2 (DC Vaccination) describing how CP abilities show a marked difference among DC subsets and antigen uptake routes, as these differences affect the results of DC vaccination.
Figure 1, antigen presentation of MHC II should be added. PPRs should be changed to PRRs.
→ We have corrected “PPRs” to “PRRs.”
Lines 197-200, the description is confused.
→ We have removed the overlapping sentence to ensure clarity.
Figure 2, ‘incorrect disulfide bods’ should be changed to ‘incorrect disulfide bonds’.
→ We have corrected “bods” to “bonds.”
There are many spelling errors.
Line 80, ‘PPRs’ should be changed to ‘PRRs’.
→ We have corrected “PPRs” to “PRRs.”
Line 103, [31,34] should move to the end of this sentence.
→ We have moved the reference [31,34] (now [35,38]) to the end of the phrase.
Line 130, -glucans should be corrected.
→ We have corrected “-glucans” to “-glucan.”
Line 202, + should be superscript.
→ We have superscripted the symbol.
Lines 260 and 292, M should be corrected.
→ We have corrected “M” to “mφ.”
Line 356, ‘SEC61 and ‘ should be corrected.
→ We have removed “,” according to the reviewer's comment.
Line 442, ‘DCs’ should be changed to ‘DCs.'.
→ We have corrected “DCs” to “DCs.”.
Line 460, ‘pattern recognition receptors (PRRs)’ should be changed to PRRs.
→ We have corrected “pattern recognition receptors (PRRs)” to “PRRs.”
Line 464, PPRs should be change to PRRs.
→ We have corrected “PPRs” to “PRRs.”
Reviewer 3 Report
The molecular pathway of ERAD processing in cross presentation was well described, and the potential to increase cross presentation efficiency for DC vaccination was well explained. However, DC vaccination is mainly applied to cancer and it would be better to further describe studies related to processes of cross presentation for cancer antigens or studies that have actually increased efficiency of DC vaccine against cancer.
Minor comments:
Page 3, line 130, the ? -glucans : Roman symbols are missing. page 8, line 356 SEC61 ? and ? are~~: Roman symbols are missing. page 9, line 362 Sec61 ? and a ~~: Roman symbols are missing. Page 5, line 197-200 – The same content sentence is described twice. Page 10 line 442 – activity of DCs? Cancer UPR~~.. “.” is missing between sentences. There are no titles for each Figure.
Author Response
Response to reviewers’ comments
Reviewer 3
The molecular pathway of ERAD processing in cross presentation was well described, and the potential to increase cross presentation efficiency for DC vaccination was well explained. However, DC vaccination is mainly applied to cancer and it would be better to further describe studies related to processes of cross presentation for cancer antigens or studies that have actually increased efficiency of DC vaccine against cancer.
→ We have added a paragraph discussing DC vaccination to show the results of DC vaccination for cancer therapy (section 2.2, DC Vaccination).
Minor comments:
Page 3, line 130, the ? -glucans : Roman symbols are missing. page 8, line 356 SEC61 ? and ? are~~: Roman symbols are missing.
→ We have revised "SEC61 and " to "SEC61α and β."
page 9, line 362 Sec61 ? and a ~~: Roman symbols are missing.
→ We have revised "SEC61" to "SEC61α."
Page 5, line 197-200 – The same content sentence is described twice.
→ We have deleted the second sentence.
Page 10 line 442 – activity of DCs? Cancer UPR~~.. “.” is missing between sentences. There are no titles for each Figure.
→ We have added titles for each figure as follows:
Figure 1 Maturation of endosomes and cross-presentation (CP).
Figure 2 Recognition of extracellular proteins in CP.
We have also added “.”, between sections.
